JP2010172318A - Apparatus for and method of culturing liquid spawn of mushroom - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an apparatus for and a method of culturing the liquid spawns of mushroom, capable of favorably conducting both the feed and exhaust of air inside and outside of a tank respectively in producing the spawns. <P>SOLUTION: The apparatus for culturing liquid spawns includes a culture tank 1, and aerator 2 for the culture solution in the tank 1. In the apparatus, the aerator 2 includes an air-feed pipe 3 for feeding air into the culture solution, an exhaust pipe 4 for exhausting air in the tank 1, and a by-pass pipe 5 with on/off valve 22 for by-passing the air-feed pipe 3 and the exhaust pipe 4 out of the tank 1. The air-feed pipe 3 includes: a first valve 10 to block the inflow of air in making a spawn inoculation; a second valve 12 to block the backflow of the culture solution in making a heat sterilization; and a filter 11 to clean in-flow air. Besides, the exhaust pipe 4 includes: a manometer 16 installed detachably; and a third valve 17 and a fourth valve 18 installed on both sides of the manometer 16 with it therebetween and to be closed when the manometer is detached. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a liquid inoculum culture apparatus for cultivating an inoculum (liquid inoculum) using a liquid as a medium base material and a method for culturing a liquid inoculum of a mushroom.

  Inoculum is used in the artificial cultivation of mushrooms such as enoki mushrooms, bean mushrooms, maitake mushrooms, nameko, and eringi. The seed fungus is a fungus that is obtained by culturing a secondary mycelium of mushrooms in a solid medium or liquid medium based on sawdust and chips, and is like a seed when compared to a plant. Mushrooms can be harvested by inoculating this inoculum into the medium and passing through the manufacturing process (culture process, generation process, breeding process) of mushrooms (fruit bodies).

  There are two methods for producing inoculum, solid culture and liquid culture, but solid culture is common. However, solid culture is more complicated and time consuming than liquid culture. Therefore, the space for culture is also reduced accordingly. In addition, since the inoculum is cultured using a culture bottle, it is necessary to prepare a large number of culture bottles, and handling of the culture bottle becomes complicated.

  In addition, in the case of solid culture, since it is impossible to confirm the quality of the inoculum unless the inoculum is actually inoculated into the medium and the mushrooms are cultivated, a cultivation loss occurs when there is a problem with the inoculum. There is also a problem that it takes a long time to recover even when recovering.

  Thus, liquid culture has attracted attention. For example, Patent Document 1 discloses a method for producing a liquid inoculum of a mushroom. FIG. 7 schematically shows the steps of the method for culturing the liquid inoculum of this mushroom. In addition, the white circle in FIG. 7 shows the state which has opened the valve, and the black circle shows the state which has closed the valve.

JP 2002-51639 A

  In the heat sterilization process, since the air in the culture tank expands due to heating, the air must be exhausted to the outside of the culture tank. According to Patent Document 1, as shown in (A) in the upper part of FIG. When exhaust is performed by opening the valve (52) in the heat sterilization process (broken arrows indicate the flow of exhaust), a filter is provided in the pipe on the valve (52) side when air flows in the cooling process. Because it is not, germs also invade with it. Further, as shown in (B) in the lower part of FIG. 7, when sterilization is performed by opening the valves (45) and (48) in the heat sterilization process (the broken line arrow indicates the flow of exhaust), it flows into the cooling process. Since the air to be passed through the filter (44), the inside of the culture tank is not contaminated. However, since the pressure in the culture tank increases during heat sterilization, the culture solution flows backward to the outside of the culture tank (the solid line arrow indicates the reverse flow of the liquid). Further, according to Patent Document 1, since the pressure gauge (54) cannot be removed, the pressure gauge is easily damaged during heat sterilization.

  Therefore, an object of the present invention is to provide a liquid inoculum culture apparatus and a mushroom liquid inoculum culture method capable of satisfactorily supplying and exhausting air inside and outside the tank in the production process.

  In order to achieve the above object, the present invention includes a culture tank for culturing inoculum by containing a culture solution, and an aeration device for supplying air to the culture solution in the culture tank to perform explosion. The aeration apparatus has an air supply pipe that feeds air taken in from the inlet into the culture medium in the culture tank from an outlet at the bottom of the tank, and an exhaust pipe port that supplies the air supplied to the culture liquid in the culture tank. An exhaust pipe for exhausting further out of the culture tank, and a bypass pipe with an open / close valve for bypassing the air supply pipe and the exhaust pipe outside the culture tank. A first valve that can be opened and closed on the inlet side so as to prevent the inflow of air to the inlet, and at the downstream side of the first valve and downstream of the branch portion of the bypass pipe, Inflow from an inflow port interposed between a second valve that is openable and closable so as to prevent the culture medium in the culture tank from flowing backward, and the first valve and a branch portion of the bypass pipe on the downstream side. The exhaust pipe is provided with a pressure gauge that is detachably installed on the downstream side of the branch of the bypass pipe, and the pressure gauge that is disposed on both sides of the pressure gauge is removed. A mushroom liquid inoculum culture apparatus comprising a third valve and a fourth valve that are sometimes closed.

  Using such a mushroom liquid inoculum culture device, a heat sterilization preparation step performed before heat sterilization of the culture solution, a heat sterilization step of heat sterilizing the culture solution in the culture tank in a high temperature atmosphere, The liquid inoculum of mushrooms can be cultured by sequentially performing a cooling step of cooling, an inoculation step of inoculating the culture solution with an inoculum, and an inoculum culture step of culturing the inoculum.

  That is, in the heat sterilization preparation step, the culture solution is stored in the culture tank, the pressure gauge detachably fixed to the attachment port of the exhaust pipe is removed, and the lid is closed on the attachment port. By removing the pressure gauge before heat sterilization, it is possible to prevent damage to the pressure gauge due to heat. Also, by closing the lid on the pressure gauge mounting port, it is possible to prevent the inflow of contaminated air from the mounting port.

  In the heat sterilization step, the first valve and the open / close valve are opened, the second valve, the third valve and the fourth valve are closed, and the expanded air in the culture tank is passed from the exhaust pipe, through the bypass pipe and the air supply pipe, It can be discharged from the inlet of the air pipe. It is possible to prevent the pressure in the culture tank from becoming too high due to the expansion of the air in the culture tank accompanying heating. In addition, the second valve in the closed state can prevent the culture solution from flowing back out of the culture tank from the inside of the culture tank in a high pressure state.

  In the cooling step, with the third valve and the fourth valve closed and the exhaust pipe closed, the lid is removed and a pressure gauge is attached to the attachment port, and the first valve and the open / close valve are opened, and the second valve, the third valve With the valve and the fourth valve closed, the inflow of air into the culture tank due to the condensation of the air in the culture tank due to cooling from the inlet of the air supply pipe, the air supply pipe, the filter, the bypass pipe, and the exhaust This is done through an exhaust pipe opening that opens into the culture tank through the tube. Since the 3rd valve and the 4th valve are closed when attaching a pressure gauge, the contamination by the inflow of air from the attachment port at the time of removing a lid can be suppressed to the minimum. In addition, when the air flows in, since the air pipe is provided with a filter, the air flowing from the inlet can be purified. Therefore, contamination in the culture tank can be prevented. Further, a water jacket may be provided on the outer periphery of the culture tank, and water may be circulated in the water jacket in the cooling step. Compared to natural cooling, the cooling time can be significantly shortened.

  In the inoculum inoculation step, the inoculum is inoculated into the culture solution in the culture tank. The closed first valve can prevent air from flowing into the culture tank during inoculation.

  In the inoculum culture process, the first valve, the second valve, the third valve, and the fourth valve are opened and the open / close valve is closed, and the culture solution is aerated by sending air from the inlet through the air supply pipe and from the outlet. The air in the culture tank is taken from the exhaust pipe port and exhausted from the exhaust port through the exhaust pipe.

  Moreover, it is preferable to provide a stirring device for stirring the culture solution stored in the culture tank. The stirring device may include a stirring blade and a drive motor connected to the blade shaft of the stirring blade, the upper end of the blade shaft is exposed to the outside of the tank, and the drive motor may be detachably provided on the blade shaft. . By making the drive motor detachable, the drive motor can be prevented from being damaged during heat sterilization.

  According to the present invention, a plurality of valves are provided at predetermined positions of an aeration apparatus for supplying and exhausting air in the culture tank, and by opening and closing the valves according to each step, supply of air in the culture tank and Exhaust can be performed well.

Top view of the liquid inoculum culture apparatus for mushrooms of this embodiment Partial perspective side view of the liquid inoculum culture device for mushrooms of this embodiment It is a figure which shows the attachment state of a pressure gauge or a lid | cover, Comprising: (a) is a perspective view which shows the attachment state of a lid, (b) is a perspective view which shows the attachment state of a pressure gauge. It is a figure which shows the attachment state of a pressure gauge or a lid | cover, Comprising: (a) is sectional drawing which shows the attachment state of a lid, (b) is sectional drawing which shows the attachment state of a pressure gauge The figure which shows the connection state of a drive motor and the axis | shaft of a stirring blade The figure which shows the process of the cultivation method of the liquid inoculum of the mushroom of this embodiment. The figure which shows the process of the cultivation method of the conventional liquid inoculum of a mushroom The perspective view which shows the washing stand apparatus of the liquid inoculum culture device of the mushroom of this embodiment The side view which shows the washing stand apparatus of the liquid inoculum culture device of the mushroom of this embodiment

  Embodiments of the present invention will be described below. FIG. 1 is a plan view of a liquid inoculum culture device for mushrooms according to the present embodiment, FIG. 2 is a partially transparent side view of the liquid inoculum culture device for mushrooms according to the present embodiment, and FIG. (A) is a perspective view showing the attachment state of the lid, (b) is a perspective view showing the attachment state of the pressure gauge, and FIG. 4 is a view showing the attachment state of the pressure gauge or the lid, (a) FIG. 5 is a cross-sectional view showing the state of attachment of the lid, FIG. 5B is a cross-sectional view showing the state of attachment of the pressure gauge, and FIG.

  As shown in FIGS. 1 and 2, the liquid inoculum culture apparatus in this embodiment includes a culture tank 1 for containing a culture solution and cultivating the inoculum, and air in the culture solution in the culture tank 1. An aeration apparatus 2 for supplying and performing explosion.

  The aeration apparatus 2 includes an air supply pipe 3 that sends air into the culture solution in the culture tank 1, an exhaust pipe 4 that exhausts the air supplied to the culture tank 1 to the outside of the culture tank 1, and an air supply pipe 3 and an exhaust pipe 4 and a bypass pipe 5 that bypasses the outside of the culture tank 1, and these are configured by connecting a plurality of metal pipes.

  The air supply pipe 3 is provided so as to communicate between the inside and outside of the culture tank 1, the inlet 6 opens to the outside of the culture tank 1, and the jet outlet 7 can be opened into the culture solution at the lower part of the culture tank 1. A branching portion to 5 is arranged outside the culture tank 1. The air supply pipe 3 includes an upstream air supply pipe 3 a from the inlet 6 to the branch portion and a downstream air supply pipe 3 b from the branch portion to the jet port 7.

  The spout 7 of the air supply pipe 3 may be the opening of the tip of the pipe constituting the air supply pipe 3, but the tip of the pipe is bent and a plurality of holes are provided on the side of the bent portion. Air is ejected from the plurality of holes (spout port 7), and aeration of the culture solution stored in the culture tank 1 can be performed efficiently.

  The upstream valve 3a is provided with a first valve 10 and a filter 11. The first valve 10 is provided on the inlet 6 side so as to be openable and closable so as to prevent the inflow of air into the culture tank 1 when inoculating the culture tank 1 with the inoculum. The filter 11 is interposed between the first valve 10 and the branching portion to the bypass pipe 5 on the downstream side thereof, and cleans the air flowing in from the inflow port 6.

  A second valve 12 is provided outside the culture tank 1 in the downstream side air supply pipe 3b. The second valve 12 is provided so as to be openable and closable so as to prevent the culture solution in the culture tank from flowing backward during the heat sterilization of the culture solution.

  The exhaust pipe 4 is provided in communication with the inside and outside of the culture tank 1, the exhaust pipe port 14 opens into the culture tank 1, the exhaust port 15 opens out of the culture tank 1, and the branch to the bypass pipe 5 is cultured. It is arranged outside the tank 1. The exhaust pipe port 14 is located in the upper part of the culture tank 1 and is not immersed in the culture solution. The air in the culture tank 1 can be taken in from the exhaust pipe port 14 and can be exhausted out of the culture tank 1 from the exhaust port 15. The exhaust pipe 4 includes an upstream exhaust pipe 4 a from the exhaust pipe port 14 to the branch portion and a downstream exhaust pipe 4 b from the branch portion to the exhaust port 15.

  A pressure gauge 16 for measuring the pressure in the culture tank 1 is detachably provided in the downstream exhaust pipe 4b, and a third valve 17 and a fourth valve 18 are disposed on both sides of the pressure gauge 16 therebetween. . The fourth valve 18 is provided on the downstream side (side closer to the exhaust port 15). The third valve 17 and the fourth valve 18 close the downstream exhaust pipe 4 b when the pressure gauge 16 is removed, and prevent outside air from flowing in from the attachment port 20 of the pressure gauge 16.

  As shown in FIGS. 3 and 4, the pressure gauge 16 is detachably provided by a ferrule clamp 19 at an attachment port 20 of a pipe forming the downstream side exhaust pipe 4 b. The attachment port 20 is branched and formed so as to protrude upward from the pipe, and the opening is used as the attachment port 20. The tip of the attachment port 20 is formed in a flange shape. The lower end of the pressure gauge 16 is also formed in a flange shape. The flange portion 20a of the mounting port 20 and the flange portion 16b of the pressure gauge 16 are overlapped, and the overlapped portion is surrounded by a ferrule 19a and fixed by tightening with a clamp 19b. In addition, a plug 21, which is a separate member capable of covering the attachment port 20, can be detachably attached to the attachment port 20 by a ferrule clamp 19. The lower portion of the plug 21 is formed in a flange shape. The flange portion can be overlapped with the flange portion of the attachment port 20, and the overlapped portion can be fastened and fixed by the ferrule clamp 19.

  The bypass pipe 5 connects the branch portion of the air supply pipe 3 and the branch portion of the exhaust pipe 4. The bypass pipe 5 is provided with an open / close valve 22 that opens and closes the bypass pipe 5.

  As the above-described valves 10, 12, 17, 18 and 22, needle valves are used for the first valve 10 and the fourth valve 18, and diaphragm valves are used for the second valve 12, the third valve 17 and the opening / closing valve 22. Although used, it is not limited to this.

  The culture tank 1 is formed in a sealed container with a bottom using a stainless steel plate. In addition, it is possible to use materials other than a stainless steel plate, and it is also possible to use materials, such as a plastic, not only a metal plate. Although the size of the culture tank 1 can be designed as appropriate, in this embodiment, the culture tank 1 having a diameter of 77 cm and a height of 130 cm is used. In the upper part of the culture tank 1, an opening (not shown) serving as a culture medium inlet and a lid 24 for closing the opening are provided. The lid 24 can be opened and closed and sealed with a screw member 25. A stirring device attaching portion 26 for fixing the stirring device 34 is provided at the center of the lid 24.

  In the upper part of the culture tank 1, an inoculation port 27 for introducing the adjusted culture solution adjusted in the pre-culture stage is provided. The inoculation port 27 is connected to an injection tube 29 that communicates with the adjusted culture solution in the container (conical flask) 28 containing the adjusted culture solution in the inoculum inoculation step described later.

  A discharge port 30 for discharging the culture solution in the culture tank 1 is provided at the bottom of the culture tank 1. An inspection window 31 for confirming the state of the culture solution in the culture tank 1 is provided on the side surface and the upper surface of the culture tank 1. In addition, a caster 32 is provided in the lower part of the culture tank 1 so that the culture tank 1 can be easily moved.

  A water jacket 33 serving as a cooling water flow path is provided so as to cover the outer periphery of the culture tank 1. The water jacket 33 is provided with a second outer wall on the outer periphery of the outer peripheral wall of the culture tank 1 to form a double wall structure, and cooling water can flow through the space between them. The water jacket 33 is provided in a C-shape so as to cover most of the side surface of the culture tank 1 while avoiding the observation window 31 on the side surface of the culture tank 1 and the caster 32 below the culture tank 1. The water jacket 33 has a structure in which cooling water is injected from the lower inlet 33a, and excess water flows out from the upper outlet 33b. By circulating the cooling water in the water jacket 33, heat exchange is performed between the cooling water and the culture solution in the culture tank 1, and the culture solution can be cooled in a short time.

  A stirring device 34 for stirring the culture solution in the culture tank 1 is provided. The stirring device 34 includes a stirring blade 35 and a drive motor 36. The blade portion of the stirring blade 35 is disposed inside the culture tank 1 and can stir the culture solution stored in the culture tank 1. The upper end portion of the blade shaft 35 a of the stirring blade 35 is inserted through the stirring device mounting portion 26 and protrudes out of the culture tank 1. The blade shaft 35a of the stirring blade 35 is fixed to the stirring device mounting portion 26 so as to be rotatable. Further, as shown in FIG. 5, the blade shaft 35a of the stirring blade 35 is inserted into the shaft 36a of the drive motor 36, and is fixed to the respective insertion holes 35b, 36b with screws 37 and nuts 38. The shaft 35a and the drive motor 36 are detachably fixed. With this configuration, the drive motor 36 of the stirring device 34 can be removed during heat sterilization.

  Next, the cleaning stand device 41 of the liquid inoculum culture device will be described. Although the culture tank 1 can be used repeatedly, it is necessary to wash the culture tank 1 after use. In the washing operation, the screw member 25 of the culture tank 1 is removed, the lid 24 is opened, washing water such as water is injected from the opening to wash the inner surface of the culture tank 1 and drain from the discharge port 30 below the culture tank 1. By doing. At this time, since the liquid inoculum culture device is heavy, it is difficult to perform drainage work from the discharge port 30.

  Therefore, as shown in FIGS. 8 and 9, the cleaning table device 41 includes a table 42 on which the liquid inoculum culture device 1 is placed, and an inclination means 43 that inclines the table 42. The liquid inoculum culture device 1 on the gantry 42 is tilted by the tilting means 43, and the drainage work from the discharge port can be easily performed.

  The gantry 42 includes a pedestal 44 and a fence 45 erected from the pedestal 44 so as to surround the periphery of the liquid inoculum culture device 1. The fence 45 regulates its periphery so that it does not fall off when the liquid inoculum culture device 1 is tilted. The front fence 45a is provided so as to be freely opened and closed, and the liquid inoculum culture apparatus 1 can be taken in and out by opening the front fence 45a.

  The tilting means 43 includes a support portion 46 that supports the pedestal 44 of the gantry 42 and an elevating means 47 such as a cylinder. The support portion 46 rotatably supports the front side surface of the pedestal 44 of the gantry 42 by the shaft portion 46b so that the gantry 42 can be tilted forward. The cylinder 47 connects the rear end of the support portion 46 and the rear end of the fence 45 of the gantry 42 vertically. By extending and retracting the cylinder 47 and raising and lowering the rear side of the fence 45 of the gantry 42, the gantry 42 can be tilted or returned.

[Manufacturing process of liquid inoculum]
Hereinafter, the steps for producing the liquid inoculum will be described in order. The liquid inoculum is produced through a pre-culture stage and a main culture stage.

  The culture solution used in the liquid inoculum culturing apparatus is not limited to this. For example, soy flour, hydrogen phosphate, sulfate, and saccharide are mixed well in water as nutrients. Is a culture solution. More specifically, defatted soybean powder is used as the soybean powder. The content of the defatted soybean powder in the culture solution is preferably in the range of 0.5 to 7.5 g / L, more preferably in the range of 1.5 to 4 g / L. As the hydrogen phosphate, potassium dihydrogen phosphate is used. The content of potassium dihydrogen phosphate in the culture solution is preferably in the range of 0.05 to 2.5 g / L, more preferably in the range of 0.2 to 2.5 g / L. Magnesium sulfate heptahydrate is used as the sulfate. The content of magnesium sulfate heptahydrate in the culture solution is preferably in the range of 0.05 to 2.5 g / L, more preferably in the range of 0.2 to 2.5 g / L. Refined sugar is used as the saccharide. The refined sugar is preferably in the range of 3 to 20 g / L, more preferably in the range of 6 to 19 g / L.

1. Pre-culture stage First, a strain is inoculated into a petri dish solid medium (general agar medium) from a test tube containing a preserved strain of mushrooms and cultured in a greenhouse (incubator 20 ° C.). After the mycelium spreads in the medium, 2 to 10 lumps of about 5 mm square block are taken out from the petri dish and inoculated into an Erlenmeyer flask 28 containing 500 to 1000 ml of the above culture solution. The Erlenmeyer flask 28 containing the culture solution is sterilized in advance at 121 ° C. for 15 to 20 minutes by high-pressure sterilization using an autoclave.

  Next, the Erlenmeyer flask 28 inoculated with the lump is statically cultured for one day in a room adjusted to a room temperature of about 20 ° C to 25 ° C. After stationary culture, the cells are cultured with shaking (rotation) for 6 to 11 days in an environment of 20 to 25 ° C. For shake (rotation) culture, a shake incubator was used and shaken at a rotation speed of 100 rpm, but the rotation speed of the shake incubator may be appropriately adjusted at about 50 to 150 rpm.

  As the shaking culture approaches completion, the mycelium extends into the culture. When this culture solution is used for main culture, the mycelium should be finely divided with a homogenizer or a magnetic stirrer. In addition, according to the rotation culture using a magnetic stirrer, the mycelium becomes fine as a result. This is because fine bacteria speeds up the bacteria around the main culture. The pre-culture stage is completed in about 7 to 14 days.

2. Main culture stage Next, the main culture stage is started. FIG. 6 is a diagram showing the steps of the method for culturing a liquid inoculum of a mushroom according to this embodiment. In addition, the white circle in FIG. 6 shows the state which has opened the valve, and the black circle shows the state which has closed the valve.

  In the main culture, a liquid inoculum culture apparatus in which about 360 L (or 470 L) of the culture solution is accommodated is used. In the main culture stage, A. A heat sterilization preparation step; A heat sterilization step; A cooling step; Inoculation step; The inoculum culture step is sequentially performed.

<A. Heat sterilization preparation process>
The heat sterilization preparation step is a step performed before the heat sterilization step. The clamp 19b is loosened and the pressure gauge 16 is removed. Then, the stopper 21 is fastened and fixed to the attachment port 20 with a ferrule clamp 19. Moreover, the drive motor 36 of the stirring apparatus 34 which stirs the culture solution in the culture tank 1 is removed. It is possible to prevent the pressure gauge 16 and the drive motor 36 of the agitating device 34 from being damaged due to heating in the heat sterilization process or internal entry of steam.

<B. Heat sterilization process>
In the heat sterilization step, the liquid inoculum culture device is housed in a high-pressure heat sterilization pot conventionally used in mushroom cultivation, and the entire liquid inoculum culture device is heat sterilized. In this embodiment, although it sterilizes at 120 degreeC for about 20 minutes, it is not limited to this.

  At this time, the first valve 10 and the opening / closing valve 22 are opened, and the second valve 12, the third valve 17 and the fourth valve 18 are closed and placed at a high temperature. The air in the culture tank 1 expands and the pressure in the culture tank 1 becomes high, but the expanded air in the culture tank 1 is closed because the second valve 12, the third valve 17, and the fourth valve 18 are closed. The exhaust pipe 4 is discharged from the inlet 6 of the air supply pipe 3 through the upstream exhaust pipe 4a, the bypass pipe 5 and the upstream air supply pipe 3a. It is possible to prevent the pressure in the culture tank 1 from becoming too high. Moreover, the filter 11 located in the upstream air supply pipe 3a can be sterilized with the high-temperature exhaust.

<C. Cooling process>
Next, as a preparatory step before moving to the cooling process, the first valve is temporarily closed to prevent the inflow of air from the air supply pipe 3 into the culture tank 1, and the third valve 17 and the fourth valve 18. Is closed and the inflow of air from the exhaust pipe 4 is prevented, the liquid inoculum culture apparatus after heat sterilization is taken out of the high-pressure heat sterilization pot and moved to the culture chamber.

  Then, with the third valve 17 and the fourth valve 18 closed, the clamp 19 b is loosened and the plug 21 is removed, and then the pressure gauge 16 is attached to the attachment port 20 using the ferrule clamp 19. At this time, since the third valve 17 and the fourth valve 18 are closed, it is possible to minimize contamination air from flowing into the culture tank 1 from the attachment port 20 when the stopper 21 is removed.

  Further, a clean air supply line 39 for sending clean (sterile) air is connected to the inflow port 6, and an exhaust line 40 is connected to the exhaust port 15. The clean air supply line 39 includes a purifying means (not shown) such as a filter that primarily purifies air and an air blowing means (not shown). In the present embodiment, the air blowing means is an air generator in a compressor and can be fed at a high pressure. The exhaust line 40 exhausts the outside of the culture chamber.

  In addition, the drive motor 36 of the stirring device 34 is attached to the shaft of the stirring blade 35 and the stirring device 34 is installed.

  Next, the process proceeds to a cooling step, in which the first valve 10 and the opening / closing valve 22 are opened, and the second valve 12, the third valve 17 and the fourth valve 18 are closed. In the cooling process, cooling water is injected from the lower inlet 33 a of the water jacket 33, excess water is drained from the upper outlet 33 b, and the cooling water is circulated in the water jacket 33. Heat exchange is performed between the cooling water and the culture solution in the culture tank 1. The temperature can be lowered to about 20 degrees at which the inoculum can be inoculated, and it can be performed in about 15 hours in the conventional natural cooling, which takes nearly 40 hours, and the cooling time can be shortened.

  During cooling, the air in the culture tank 1 is condensed. Along with this, the pressure in the culture tank 1 decreases, so it is necessary to flow air into the culture tank 1. This inflow of air is performed from the exhaust pipe port of the exhaust pipe 4 through the pipe in which the valve is open, that is, from the inlet 6 of the air feed pipe 3 through the upstream air feed pipe 3a, the bypass pipe and the upstream exhaust pipe 4a. . Air is forcibly fed into the culture tank 1 from the clean air supply line 39 connected to the inlet of the air supply pipe 3, and the pressure in the culture tank 1 is set to about 0.16 MPa. In addition, since the filter 11 is provided in the upstream air supply pipe 3a at the time of air inflow, the air which flows in from the inflow port 6 is purified. Therefore, contamination in the culture tank 1 can be prevented.

<D. Inoculation process>
In a state where the first valve is closed to prevent the inflow of air into the culture tank 1, the culture tank 1 is inoculated with the adjusted culture solution that has been inoculated and cultured in the pre-culture stage. The Erlenmeyer flask 28 used in the pre-culture stage is charged with one adjusted culture solution (600 ml).

  Here, a method for introducing the adjusted culture solution will be described. An injection tube 29 communicating with the conditioned culture solution in the Erlenmeyer flask 28 is connected to the inoculation port 27 at the top of the culture tank 1. A clean air supply line 41 for supplying clean (sterile) air is connected to the Erlenmeyer flask 28, and the clean air supply line 41 is connected to a purification means (not shown) such as a filter for primarily purifying the air. And air blowing means (not shown). In the present embodiment, the air blowing means is an air generator in a compressor and can be fed at a high pressure.

  Then, the pressure in the culture tank 1 is once lowered to 0.02 MPa, air is sent into the Erlenmeyer flask 28 through the clean air supply line 41, and the first valve 10 is closed to supply air to the culture tank 1. By blocking the inflow, the pressure in the Erlenmeyer flask 28 is set higher than the pressure in the culture tank 1. Using this pressure difference, the adjusted culture solution in the Erlenmeyer flask 28 can be injected into the culture tank 1. At this time, the adjusted culture solution does not come into contact with the outside air, so it is not necessary to mix various bacteria. In the past, inoculation work was performed in a clean booth, and when the culture solution was poured into the culture tank 1, it was attempted to prevent contamination with an alcohol lamp or the like. However, contamination must be completely prevented. I couldn't. However, in this embodiment, since it is directly connected to the culture tank 1, there is no fear of contamination.

<E. Inoculum culture process>
Next, culture is started. The room temperature of the culture chamber is about 22 ° C to 23 ° C. In the inoculum culture step, the first valve 10, the second valve 12, the third valve 17 and the fourth valve 18 are opened, and the open / close valve 22 is closed, so that the air sent from the inlet 6 The culture solution in the culture tank 1 is aerated by being fed from the jet outlet 7 through the downstream side air supply pipe 3b. Thus, the culture is performed while blowing air into the culture tank 1. During this aeration, since air is introduced through the filter 11, various germs do not enter the culture tank 1.

  The air supplied to the culture solution in the culture tank 1 is exhausted from the exhaust port 15 through the exhaust tube 4 through the exhaust tube port 14. By adjusting the amount of air flowing in from the inflow port 6 and the amount of air discharged from the exhaust port 15, the pressure in the culture tank 1 is set to 0.02 to 0.04 MPa.

  In addition, the culture solution is stirred by the stirring device 34 during the culture. By stirring the stirring blade 35, the inoculum can be exposed to aeration air to accelerate the growth. In addition, the mycelium of the inoculum in the culture can be finely cut by stirring with the stirring blade 35. Thus, by fragmenting the inoculum finely to prevent the colony from becoming large, it was possible to shorten the outline time from about 6 days to 3 to 4 days. Moreover, after inoculation culture | cultivation, when inoculating a culture medium in the manufacturing process of a mushroom (fruit body), clogging of the pipe | tube which conveys liquid inoculum, the nozzle for inoculation, etc. can be prevented.

  The period required for culturing in the culture tank 1 is about 3 to 6 days for enoki mushrooms and the like, and 4 to 8 days for shimeji and sea cucumbers. The progress of the culture in the culture tank 1 can be easily known by visually checking the state of the culture solution in the culture tank 1 from the inspection window 31. As the culture progresses, the mycelium extends into the culture solution in the culture tank 1, so that the culture state can be determined.

  Further, by visually recognizing the state of the culture solution in the culture tank 1 from the inspection window 31, it is possible to accurately know that the inoculum is defective or that the culture is defective. In addition, the defect of the inoculum can be easily known from the smell during culture. Thus, being able to know the quality of the inoculum prevents the bad inoculum from being used in actual cultivation, thereby preventing the occurrence of cultivation loss. This is advantageous compared to the case of using the like. Moreover, even if the inoculum is defective, it is advantageous in that it can be recovered in a short period of time by performing the steps of inoculating the sterilized and adjusted culture solution from the preparation of the culture solution again.

  Thus, a liquid inoculum can be manufactured through the above process. Mushrooms can be harvested by inoculating the liquid inoculum into the medium and passing through the manufacturing process (culture process, generation process, breeding process) of mushrooms (fruit bodies). In addition, although the growth environment and the cultivation process differ according to the kind of mushroom, the cultivation method common in each mushroom field can be employ | adopted for the cultivation method of a mushroom.

DESCRIPTION OF SYMBOLS 1 Culture tank 2 Aeration apparatus 3 Air supply pipe 3a Upstream side air supply pipe 3b Downstream side air supply pipe 4 Exhaust pipe 4a Upstream side exhaust pipe 4b Downstream side exhaust pipe 5 Bypass pipe 6 Inlet 7 Outlet 9 Air blower 10 First valve 11 Filter 12 Second valve 14 Exhaust pipe port 15 Exhaust port 16 Pressure gauge 17 Third valve 18 Fourth valve 19 Ferrule clamp 20 Mounting port 21 Plug 22 Opening / closing valve 24 Lid 27 Inoculation port 33 Water jacket 34 Stirring device 41 Washing table device

Claims (5)

A culture tank for containing the culture medium and cultivating the inoculum, and an aeration device for supplying air to the culture liquid in the culture tank and performing explosion
The aeration apparatus includes an air supply pipe that feeds air taken in from an inflow port into a culture medium in a culture tank from a jet outlet at a lower part of the tank, and an exhaust pipe port that supplies air supplied to the culture liquid in the culture tank. An exhaust pipe for exhausting more outside the culture tank, and a bypass pipe with an open / close valve for bypassing the air supply pipe and the exhaust pipe outside the culture tank,
The air supply pipe has a first valve that is openable and closable on the inlet side so as to prevent the inflow of air into the culture tank when inoculating the culture tank, and on the downstream side of the first valve. And a second valve provided on the downstream side of the branching portion of the bypass pipe so as to be openable and closable so as to prevent the culture solution in the culture tank from flowing backward during the heat sterilization of the culture solution, A filter that is interposed between the branch portion of the bypass pipe on the downstream side and purifies the air flowing in from the inflow port,
The exhaust pipe includes a pressure gauge that is detachably installed downstream of the branch portion of the bypass pipe, a third valve that is disposed on both sides of the pressure gauge and closes when the pressure gauge is removed, and a third valve A mushroom liquid inoculum culture device comprising four valves. The liquid inoculum culture apparatus for mushrooms according to claim 1, wherein a water jacket is provided on an outer periphery of the culture tank. A stirrer that stirs the culture solution stored in the culture tank is provided, the stirrer includes a stirring blade and a drive motor connected to the blade shaft of the stirring blade, and the upper end of the blade shaft is The liquid inoculum culture apparatus for mushrooms according to claim 1 or 2, wherein the apparatus is exposed to the outside of the tank, and the drive motor is detachably provided on the blade shaft. A heat sterilization preparation step performed before the heat sterilization of the culture liquid using the liquid inoculum culture apparatus according to any one of claims 1 to 3, and a heat sterilization of the culture liquid in the culture tank in a high temperature atmosphere A method of cultivating a liquid inoculum that sequentially performs a heat sterilization step, a cooling step for cooling the culture solution, a seed inoculation step for inoculating the culture solution with an inoculum, and an inoculum culture step for culturing the inoculum. ,
In the heat sterilization preparation step, the culture solution is stored in the culture tank, and after removing the pressure gauge from the attachment port of the exhaust pipe, the attachment port is closed with a lid,
In the heat sterilization step, the first valve and the open / close valve are opened, the second valve, the third valve, and the fourth valve are closed, and the expanded air in the culture tank is passed from the exhaust pipe through the bypass pipe and the air supply pipe. , While discharging from the inlet of the air pipe, the second valve in the closed state prevents the culture fluid in the culture tank from flowing backward,
In the cooling step, with the third valve and the fourth valve closed and the exhaust pipe closed, the lid is removed, a pressure gauge is attached to the attachment port, the first valve and the open / close valve are opened, the second valve, With the third valve and the fourth valve closed, the inflow of air into the culture tank due to the condensation of the air in the culture tank due to cooling from the inlet of the air supply pipe, the air supply pipe, the filter, and the bypass From the exhaust pipe opening that opens into the culture tank through the pipe and the exhaust pipe,
In the seed inoculation step, inoculate the culture tank with the inoculum in a state where the first valve is closed to prevent the inflow of air into the culture tank,
In the inoculum culture step, the first valve, the second valve, the third valve, and the fourth valve are opened and the open / close valve is closed, and the air from the inflow port is fed into the culture tank through the air supply pipe into the culture tank. A method for cultivating a liquid inoculum of a mushroom, characterized by performing aeration of liquid and exhausting air in a culture tank from an exhaust pipe through an exhaust pipe through the exhaust pipe. The method for cultivating a liquid inoculum of a mushroom according to claim 4, wherein water is circulated in a water jacket in the cooling step.