Pouch storage device and microbial culture device

Abstract

To provide a pouch containment device that facilitates the handling of pouches when culturing microorganisms using pouches, and a microbial culture device that prevents microorganisms and contaminants from entering the pouches or culture tanks. [Solution] The pouch storage device (10) of the present invention has a skeletal structure composed only of the outline and outer circumference of the central part in the height direction of a hypothetical container that surrounds the pouch (1). The microbial culture device (100) of the present invention has the pouch storage device (10).

Description

Technical Field

[0001] The present invention relates to a technique for culturing microorganisms such as lactic acid bacteria using a culture tank.

Background Art

[0002] Techniques for culturing microorganisms using a culture tank are well-known. An example of such a culturing technique will be described with reference to FIGS. 20 and 21. As shown in FIG. 20, first, a culture medium A is accommodated in a Erlenmeyer flask 31, stoppered with a stopper 31A, and sterilized in an autoclave 32. Then, the microorganism to be cultured is inoculated into the culture medium A in the Erlenmeyer flask 31 and cultured. After culturing the microorganism in the Erlenmeyer flask 31, as shown in FIG. 21, the culture medium (the culture medium in which the microorganism has been cultured) in the Erlenmeyer flask 31 is supplied into the culture tank 33 through the inoculation port 33A of the culture tank 33 (hereinafter, in this specification, supplying a microorganism into a pouch or a culture tank is described as "inoculation" and is distinguished from the supply of air or steam).

[0003] However, as shown in FIG. 21, when inoculating the culture medium A in the Erlenmeyer flask 31 into the culture tank 33 through the inoculation port 33A of the culture tank 33, the Erlenmeyer flask 31 is opened and the inoculation port 33A of the tank 33 is opened. Therefore, there is a possibility that miscellaneous bacteria, other microorganisms, and contaminants may enter from the opened portion, contaminating the culture medium in the Erlenmeyer flask 31 or growing in the culture tank 33 (so-called "contamination" problem). In particular, in the culture of microorganisms (e.g., lactic acid bacteria) contained in food and drink, the above-mentioned "contamination" problem causes great damage, so the possibility must be minimized as much as possible. Also, in order to prevent contamination, for example, there are methods such as spraying sterilizing alcohol at the time of opening or using a flame near the inoculation port 33A, but there is a possibility that the operator may be burned or injured. Furthermore, performing operations that may cause problems such as contamination, burns, and injuries places a physical or mental burden on the operator.

[0004] Other conventional technologies include cultivation techniques using pouches with a spout (see, for example, Patent Document 1). However, since the pouch is made of a synthetic resin bag, it is difficult to maintain a consistent shape even when filled with culture medium, for example, when the spout is facing downwards. Therefore, it presents a problem in that it is difficult to handle when inoculating into a culture tank. Furthermore, after culturing the necessary microorganisms, when inoculating the cultured microorganisms and culture medium into the culture tank, the spout of the pouch and the inlet of the culture tank are opened, which still presents the problem of other microorganisms and contaminants entering (the so-called "contamination problem"). [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] Japanese Patent Application Publication No. 8-266268 [Overview of the project] [Problems that the invention aims to solve]

[0006] This invention was proposed in view of the problems of the prior art described above, and aims to provide a pouch containment device that facilitates the handling of pouches when culturing microorganisms to be cultured using pouches, and a microbial culture device that can prevent microorganisms and contaminants from entering the pouch or culture tank. [Means for solving the problem]

[0007] The pouch storage device of the present invention (10: Pouch storage device of claim 1) is characterized in that it has a skeletal structure composed only of the outline and outer circumference of the central part in the height direction of a hypothetical container that surrounds the pouch (1).

[0008] Here, the aforementioned skeletal structure is The contours of the upper surface (11) and the lower surface (12) formed in a fan shape, Two first straight lines (15) connect the portions of the two radii (13) of the sectors of the upper surface (11) and the lower surface (12) that are closer to the arc than the center, Preferably, the sectors of the upper surface (11) and lower surface (12) include two second straight lines (16) that connect the positions opposite to the starting point of the first straight line (15) in each of the circular arcs (14) of the sector. Furthermore, it is preferable that the central angle of the sector is 120°.

[0009] The microbial culture apparatus (100) of the present invention is characterized by having the pouch containment device (10: the pouch containment device according to either claim 1 or 2). In this case, it is preferable that the microbial culture apparatus (100) of the present invention allows the inoculation port (33A) of the culture tank (33) to be connected to the pouch storage device (10) while the inoculation port (33A) of the culture tank (33) is not open. Furthermore, the microbial culture apparatus (100) of the present invention is preferably configured in combination with a retort sterilizer or autoclave (32) for sterilizing the pouch (1). When used in combination with an autoclave (32), it is preferable to provide a vent filter (35) at the spout (1A) of the pouch (1). Alternatively, if the autoclave (32) is not provided with a vent filter (35), a container (36) is provided inside the autoclave (32) that is open at the top, filled with water, and capable of filling the pouch (1) while it is immersed in water (W). Preferably, the amount of water (W) filled in the container (36) is such that the water level (H2) (inside the container) when the pouch (1) is immersed is 30% or more of the liquid level (H1) of the liquid culture medium inside the pouch (1).

[0010] The microbial culture apparatus (100) of the present invention has an inoculation line (40) that connects the spout (1A) of the pouch (1) and the inoculation port (33A) of the culture tank (33), and the inoculation line (40) is A connection port (41) connected to a sterilization steam supply source (47) and a compressed air (sterilized air that has passed through a 0.2 μm filter) supply source (48), A tank connection port (42) connected to the inoculation port (33A) of the culture tank (33), It has outlets (43) for sterilizing steam and compressed air, It has a piping system (44) that connects to the pouch (1) side via a branch section (40A), The piping system (44) connected to the pouch (1) is preferably connected to a pouch connection pipe (45: hand valve interposed piping), and the pouch connection pipe (45) preferably has a pouch connection portion (45A) that can be connected to the spout (1A) of the pouch (1) and has a plurality (for example, two) on-off valves (46-1, 46-2) interposed therein.

[0011] The microbial culture method using the microbial culture apparatus (100) of the present invention is: The process includes a step of placing a pouch (1) having a spout (1A) into a pouch storage device (10), the pouch storage device (10) having a skeletal structure composed only of the outline and outer circumference of the central part in the height direction of a hypothetical container that surrounds the pouch (1), A process of sterilization using a retort sterilizer or autoclave (32), The process involves culturing microorganisms (e.g., lactic acid bacteria) in a sterilized pouch (1), The process includes a step of transferring cultured microorganisms and culture medium from a pouch (1) into a culture tank (33) via an inoculation line (40).

[0012] When sterilizing in an autoclave (32), it is preferable to install a vent filter (35) in the spout (1A) of the pouch (1) and then perform the sterilization process. Alternatively, if sterilization is performed in an autoclave (32) without a vent filter (35), it is preferable to immerse 30% or more of the pouch (1) in water before performing the sterilization process. [Effects of the Invention]

[0013] According to the pouch accommodating device (10) or the microorganism culturing device (100) of the present invention having the above-described configuration, since the pouch (1) can be accommodated in the pouch accommodating device (10), even when the spout (1A) faces downward and the pouch alone cannot maintain a certain shape, the framework-like pouch accommodating device (10) composed only of the contour line of the container maintains a certain shape. Therefore, the pouch (1) can be moved together with the pouch accommodating device (10) and connected to other members (for example, the inoculation line 40). Here, the framework-like pouch accommodating device (10) composed only of the contour line of the container is lightweight, and when handling the pouch accommodating device (10) with the pouch (1) accommodated therein, it does not impose a great burden on the operator as compared with handling the pouch alone.

[0014] If the framework-like form of the pouch accommodating device (10) has the contours of the upper surface (11) and the bottom surface (12) formed in a fan shape, a plurality of adjacent pouch accommodating devices (10) can be arranged adjacent to each other so that the planar shapes of the fan shapes of the upper and lower surfaces of the plurality of pouch accommodating devices (10) become circular. Autoclaves and retort sterilizers are often circular, and in such a state, the storage efficiency is improved and the heating state during sterilization becomes uniform. For example, if the central angle of the fan shape is 120°, by arranging three pouch accommodating devices (10) adjacent to each other, the planar shapes of the upper surface (11) and the bottom surface (12) become circular, and the three pouch accommodating devices (10) are in a stable state.

[0015] Further, if the microorganism culturing device (100) of the present invention is configured in combination with a retort sterilizer or an autoclave (32) for sterilizing the pouch (1), the operation of sterilizing the pouch (1) is efficiently performed. Here, when combined with a retort sterilizer, since the retort sterilizer is provided with a pressurizing mechanism and a cooling mechanism, even if the pouch (1) is heated during sterilization, the medium in the pouch (1) does not boil, so it is possible to prevent the pouch (1) from expanding and bursting. On the one hand, since the autoclave (32) is not provided with a pressurization mechanism or a cooling mechanism, when heating in the autoclave (32) during the sterilization of the pouch (1), the medium in the pouch (1) may boil and expand, causing the pouch to rupture. However, when combined with the autoclave (32), if a vent filter (35) is provided at the spout (1A) of the pouch (1), the pressure adjustment function of the vent filter (35) can prevent the pouch (1) from rupturing. Alternatively, when combined with the autoclave (32), a container (36) is provided in the autoclave (32) that is open at the top, filled with water, and capable of filling the pouch (1) while immersed in the water. If the water filling amount in the container (36) is such that the water level (H2) in the container (36) when the pouch (1) is immersed is 30% or more of the liquid medium level (H1) in the pouch (1), the rupture of the pouch (1) can also be prevented. This has been confirmed by the inventor's experiments.

[0016] In the present invention, if there is an inoculation line (40) connecting the spout (1A) of the pouch (1) and the inoculation port (33A) of the culture tank (33), after culturing microorganisms in the pouch (1), connect the spout (1A) of the pouch (1) and the inoculation line (40), and connect the inoculation port (33A) of the culture tank (33) and the inoculation line (40), then the medium (medium in which microorganisms have been cultured) in the pouch (1) can be inoculated into the culture tank (33) via the inoculation line (40). Here, if the spout (1A) of the pouch (1) and the inoculation line (40) are connected, compared with the case where the spout (1) of the pouch (1) and / or the inoculation port (33A) of the culture tank (33) are open, the possibility of foreign bacteria, other microorganisms, and contaminants entering the pouch (1) or the culture tank (33) is low, and the so-called "contamination" problem is extremely unlikely to occur. And by opening and closing the valve of the inoculation line (40), the medium (cultured microorganisms) in the pouch (1) can be inoculated into the culture tank (33) via the inoculation line (40).

[0017] Furthermore, if the inoculation line (40) has a connection port (41) connected to a sterilization steam supply source (47) and a compressed air supply source (48), a tank connection port (42) connected to the inoculation port (33A) of the culture tank (33), and an outlet (43) for sterilization steam and compressed air, the inoculation line (40) can be sterilized by connecting it to the sterilization steam supply source (47) and supplying sterilization steam (high-temperature steam), which is then discharged from the outlet (43). Therefore, even if bacteria enter the inoculation line (40), the bacteria are reliably removed before inoculation into the culture tank (33). Furthermore, by connecting to a compressed air supply source (48) and discharging the supplied compressed air from the outlet (43), the inoculation line (40), which has been heated by sterilizing steam (high-temperature steam), can be cooled and its interior can be dried. Therefore, there is less risk of the microorganisms cultured in the pouch (1) being killed or damaged by the high heat of the heated inoculation line (40).

[0018] Furthermore, if the system has a piping system (44) connected to the pouch (1) side via a branching section (40A), and the piping system (44) connected to the pouch (1) side is connected to a pouch connection pipe (45), and the pouch connection pipe (45) has a pouch connection section (45A) that can be connected to the spout (1A) of the pouch (1) and is equipped with a plurality (for example, two) on-off valves (46-1, 46-2), then when sterilizing the inoculation line (40), the two on-off valves (46-1, 46-2) equipped in the pouch connection pipe (45) can be closed, and the area of ​​the piping between the on-off valve (46-1) on the inoculation line side (culture tank 33 side) and the on-off valve (46-2) on the pouch side of the pouch connection pipe (45) can be left empty. In this state, even if sterilizing steam (high-temperature steam) is passed through the inoculation line (40), the unfilled area between the on / off valves (46-1, 46-2) in the pouch connection piping (45) provides strong insulation, completely preventing the high heat of the high-temperature steam from being transferred to the pouch (1). Therefore, the culture medium (inside pouch 1) containing microorganisms cultured in pouch (1), and the culture medium located in the area near the pouch-side on / off valve (46-2) of the pouch connection piping (45), does not receive heat transfer from the high-temperature steam supplied during sterilization of the inoculation line. As a result, the microorganisms cultured in pouch (1) are not exposed to the heat of the high-temperature steam flowing through the inoculation line (40) during sterilization of the inoculation line, preventing them from being killed or damaged by the high heat transfer from the steam.

[0019] The method using the microbial culture apparatus (100) of the present invention includes the step of placing a pouch (1) having a spout (1A) into a pouch storage device (10). The pouch storage device (10) has a skeletal structure composed only of the outline and outer circumference of the central part in the height direction of a hypothetical container that surrounds the pouch (1). Therefore, even if the pouch alone cannot maintain a certain shape, it can maintain a certain shape when placed in the skeletal pouch storage device (10) composed only of the outline of the container. With the pouch (1) placed in the pouch storage device (10), the pouch (1) and the pouch storage device (10) can be moved together and connected to other components (for example, an inoculation line 40). Furthermore, the method using the microbial culture apparatus (100) of the present invention includes the steps of sterilizing in a retort sterilizer or autoclave (32), culturing microorganisms in the sterilized pouch (1), and moving the cultured microorganisms and culture medium from the pouch (1) into the culture tank (33) via the inoculation line (40). Therefore, the spout (1A) of the pouch (1) and the inoculation port (33A) of the culture tank (33) can be connected, and the culture medium in the pouch (1) can be inoculated into the culture tank (33) via the inoculation line (40). Compared to the case where the spout (1) of the pouch (1) and / or the inoculation port (33A) of the culture tank (33) are open, the possibility of bacteria and other microorganisms or contaminants entering the pouch (1) when connecting the spout (1A) of the pouch (1) and the inoculation line (40) is low. Similarly, the possibility of bacteria and other microorganisms or contaminants entering the culture tank (33) is also low. Therefore, the possibility of so-called "contamination" problems occurring is extremely low. [Brief explanation of the drawing]

[0020] [Figure 1] This is a front view showing an embodiment of the microbial culture apparatus of the present invention. [Figure 2] This is a side view showing an embodiment of the microbial culture apparatus of the present invention. [Figure 3] This is a plan view showing an embodiment of the microbial culture apparatus of the present invention. [Figure 4] This is a front view showing the pouch containment device for the microbial culture apparatus of the present invention. [Figure 5] This is a bottom view of the pouch storage device. [Figure 6] This is an explanatory bottom view showing the pouch storage device with a handle attached to its bottom surface. [Figure 7] This is an explanatory front view showing the pouch in the pouch storage device. [Figure 8] This is an explanatory bottom view showing the pouch in the pouch storage device. [Figure 9] This is an explanatory bottom view showing how the spout of the pouch is attached to the bottom surface of the pouch storage device. [Figure 10] This is an explanatory side view showing a pouch housed in the pouch housing device, with the pouch connection piping connected to the spout of the pouch. [Figure 11] This is an explanatory plan view showing the state in which three pouches are stored in the pouch storage device. [Figure 12] This is an enlarged front view showing the pouch storage device with the transport chain attached. [Figure 13] This is an explanatory side view illustrating how to transport the pouch storage device, which contains a pouch with an inoculation pipe connected to the spout. [Figure 14] This is a diagram illustrating the inoculation line connecting the pouch and the culture tank. [Figure 15] This is an explanatory diagram for the pouch. [Figure 16] This flowchart shows the procedure for a microbial culture method using the microbial culture apparatus of the present invention. [Figure 17] This is an explanatory diagram of a vent filter. [Figure 18] This is an explanatory diagram of the vent filter mounting bracket used to attach the vent filter to the spout of the pouch. [Figure 19] This is an explanatory diagram showing an example of a mechanism to prevent the pouch from bursting. [Figure 20] This is an explanatory diagram of one step in conventional microbial culture technology. [Figure 21] This is an explanatory diagram of the process that is performed after the process shown in Figure 20 in the prior art. [Modes for carrying out the invention]

[0021] Hereinafter, embodiments of the microbial culture apparatus 100 of the present invention will be described with reference to Figures 1 to 19. First, with reference to Figures 1 to 3, a microbial culture apparatus 100 according to the illustrated embodiment will be described. In Figures 1 to 3, the microbial culture apparatus 100 includes a pouch 1 filled with a culture medium (e.g., liquid medium) for culturing microorganisms to be cultured, a pouch storage device 10 capable of accommodating the pouch 1, a culture tank 33, and an inoculation line 40 connecting the spout 1A of the pouch 1 to the inoculation port 33A of the culture tank 33 (Figures 1 and 2). In Figures 1 and 2, the pouch 1 as it is housed in the pouch housing device 10 is shown by a dotted line. The pouch 1 will be described later with reference to Figure 15. The pouch containment device 10 has a skeletal structure composed only of the outline and outer circumference of the central part in the height direction of a hypothetical container that encloses the pouch 1 in a fitted state. The pouch containment device 10 is supported by two support rods 19 (Figures 2 and 3) via a connecting structure 37 (Figures 2 and 3) to the drive shaft housing 39 (Figures 1 and 2) on the culture tank 33 side. The specific component configuration of the pouch containment device 10 will be described in detail with reference to Figures 4 to 9. The inoculation line 40 is equipped with two on / off valves 46-1 and 46-2. The inoculation line 40 will be described in detail with reference to Figure 14.

[0022] In Figures 1 to 3, the culture tank 33 is surrounded and supported by a tank housing 34. A motor 38 is provided above the culture tank 33, and the motor 38 drives a media stirring vane (not shown) inside the culture tank 33. The drive shaft (not shown) of the motor 38 is located inside a drive shaft housing 39 (Figures 1 and 2). The area directly below the motor 38 in the tank housing 34 and the drive shaft housing 39 is connected and supported by a first support rod 51, a first bracket 53, a second support rod 52, and a second bracket 54. The first support rod 51 is fixed to the tank housing 34 side by a support portion 34A, the first bracket 53 is fixed to the drive shaft housing 39 side by a support portion 39A (Figure 1), the second support rod 52 is fixed to the tank housing 34 side by a support portion 34B, and the second bracket 54 is fixed to the drive shaft housing 39 side by a support portion 39B (Figure 1). In Figure 1, the culture tank 33 is provided with a viewing window 55, which is provided to allow the inside of the culture tank 33 to be visually inspected from outside the tank. Three observation windows 55 are provided vertically in a continuous arrangement to determine the amount of culture medium (a medium for culturing microorganisms to be cultured) to be inoculated into the culture tank 33 based on the liquid level of the medium. The number of observation windows 55 is appropriately selected according to the capacity and size of the tank.

[0023] Next, the pouch storage device 10 and pouch 1 will be described with reference to Figures 4 to 10. As shown in Figures 4 and 5, when the pouch 1 (see Figures 7 and 8) is fitted into the pouch, the pouch housing device 10 has a structure that resembles the framework of a hypothetical container (a structure consisting only of the outline of the container). The frame-like structure of the pouch storage device 10, as shown in Figures 4 and 5, includes the contours of the fan-shaped top surface 11 and bottom surface 12, two first straight lines 15, and two second straight lines 16. In Figures 4 and 5, the first straight lines 15 connect the portions of the two radii 13 of the fan-shaped top surface 11 and bottom surface 12 that are closer to the arc than the center. Each of the second straight lines 16 extends parallel to each of the two first straight lines 15, connecting the positions opposite the starting point of the first straight lines 15 in the arcs 14 of the fan-shaped top surface 11 and bottom surface 12. In Figure 5, the first and second straight lines 15 and 16 extend in a direction perpendicular to the plane of the paper, and in Figure 4, they extend in the vertical direction. In Figure 4, the straight lines 15 and 16 overlap, and only one is shown. As shown in Figure 5, the top surface 11 and the bottom surface 12 have the same configuration in plan view. The sector-shaped contours of the top surface 11 and bottom surface 12 are composed of the two radii 13 and the sector arc 14 mentioned above, as well as a central line 17 extending from the two intersection points of the radii 13 and the sector arc 14. The central angle θ in the sectors of the top surface 11 and bottom surface 12 is set to 120°.

[0024] In Figure 4, an intermediate frame 18 exists at a position midway between the top surface 11 and the bottom surface 12 in the height direction (up and down direction in Figure 4). Although not clearly shown, the outline of the intermediate frame 18 is composed of two radii 13 and a sector-shaped arc 14, similar to the top surface 11 and bottom surface 12 described above. However, the intermediate frame 18 does not include the member corresponding to the center line 17 on the bottom surface 12 and top surface 11. In Figure 4, rod connecting members 21 are positioned above the intermediate frame 18 in each of the two second straight lines 16. The rod connecting members 21 are connecting members on the pouch housing device 10 side, and these connecting members are for connecting and fixing the pouch housing device 10 to the connection structure 37 on the drive shaft housing 39 side via two support rods 19 (Figures 2 and 3) (see Figures 2 and 3). The rod connecting members 21 are also shown in Figure 5. Furthermore, in Figure 4, attachments 25 for engaging the chain of the transport device are provided at four locations on the bottom surface 12. The attachments 25 are arranged in a manner where two overlap in a direction perpendicular to the plane of the paper, so only two are shown in Figure 4. The attachment of the chain will be described later with reference to Figures 12 and 13.

[0025] Although not explicitly shown in the diagram, attachments for engaging the chain of the transport device are provided not only on the bottom surface 12 but also on the top surface 11. However, as will be described later with reference to Figures 12 and 13, when transporting the pouch storage device 10 using the chain of the transport device, the bottom surface 12 in Figure 4 is at the top and the top surface 11 is at the bottom during transport. When placing the pouch storage device 10 on a flat surface in this state, if the attachment on the top surface 11 is made to protrude vertically (up and down) in Figure 4, like the attachment 25 on the bottom surface 12, the pouch storage device 10 placed on the flat surface will become unstable. Therefore, when attaching chain engagement attachments to the top surface 11, these attachments must protrude horizontally (left and right in Figure 4), not vertically. Although not shown in Figure 5, a handle 22 for transporting the pouch storage device can be attached to the bottom surface 12 of the pouch storage device 10, as shown in Figure 6. In Figure 6, the handle 22 is attached by a mounting member 22A near the center of the center line 17 that makes up the bottom surface 12. Furthermore, as shown in Figures 5 and 6, a spout attachment 23 is provided for attaching the spout 1A of the pouch 1 when the pouch 1 is placed in the pouch storage device 10. The manner in which the pouch 1 is attached to the pouch storage device 10 will be described later with reference to Figures 7 to 9.

[0026] Figures 7 and 8 show the pouch 1 housed in the pouch housing device 10, as described in Figures 4 to 6. In Figure 7, the dotted line indicates the ridge of the pouch 1 (the inside of the sealed portion), which is folded and located on the inside of the pouch 1 (an area not visible in Figure 7). When attached to the pouch housing device 10 (see Figures 1 to 3), the pouch 1 is positioned with the spout 1A, which is the entrance and exit for the culture medium and microorganisms to be cultured, facing downwards within the pouch housing device 10. The ends of the pouch 1 opposite to the spout 1A (the upper ends in Figure 7) are held to the pouch housing device 10 (the upper surface 11 side) by the retainer 24. Pouch 1 contains a culture medium in which microorganisms to be cultured (e.g., lactic acid bacteria) have been injected, or a culture medium in which microorganisms to be cultured have been cultured, within the pouch containment device 10. Pouch 1 cannot maintain a constant shape on its own, but by being housed in the pouch containment device 10, which is a frame-like structure (composed of the aforementioned radius 13, sector-shaped arc 14, first straight line 15, second straight line 16, and center line 17), it can maintain or retain a constant shape. When pouch 1 is placed in the pouch storage device 10, it can be moved together with the pouch storage device 10 and connected to other components (for example, the inoculation line 40 shown in Figure 14). Furthermore, the pouch storage device 10, which is a frame-like structure composed only of outlines, is lightweight, and handling pouch 1 while it is stored in the pouch storage device 10 does not impose a significant burden on the worker compared to handling pouch 1 alone. Pouch 1 will be discussed later, referring to Figure 15.

[0027] As shown in Figure 8, when storing the pouch 1 in the pouch storage device 10, the spout 1A of the pouch 1 can be fixed by fitting it into the spout mounting fixture 23 (see Figures 5 and 8). The spout mounting fixture 23 is composed of an inverted U-shaped flat flange 1B (shown as a solid line in Figure 9 and a dotted line in Figure 8) provided at a distance from each other, and the spout mounting fixture 23 is provided on the bottom surface 12 of the pouch storage device 10. Figure 9 shows how the spout 1A of pouch 1 is fitted into the spout mounting fixture 23. As described above, the spout mounting fixture 23 is made up of an inverted U-shaped plate, and the spout 1A can be fixed by the spout mounting fixture 23 by fitting the inverted U-shaped plate into the gap between the two flat flanges 1B on the spout 1A side (in a tight fit). The size and shape of the inverted U-shaped plate may be arbitrarily changed to match the shape of the spout 1A of pouch 1.

[0028] As described above with reference to Figure 7, the spout 1A of the pouch 1 housed in the pouch housing device 10 faces downwards. As shown in Figure 10, the pouch connection pipe 45 is connected to the spout 1A of the pouch 1. The pouch connection piping 45 is fitted with on / off valves 46-1 and 46-2, and is connected to the spout 1A of pouch 1 via the pouch connection section 45A. The inoculation line 40, including the pouch connection pipe 45, will be described later with reference to Figure 14.

[0029] In the illustrated embodiment, as shown in Figure 11, it is possible to handle the pouch storage device 10, each containing a single pouch 1, in a combined state of three such devices. The combined pouch storage device 10 is denoted by reference numeral 10-1. The combination 10-1 of pouch storage devices 10 shown in Figure 11 is formed by combining three pouch storage devices 10 shown in Figures 4 to 9 adjacent to each other, arranging them so that the central angle θ (θ=120°) of the sectors of the top surface 11 and bottom surface 12 totals 360°. Each of the three pouch storage devices 10 (Figures 4 to 9) that make up the combination 10-1 (Figure 11) contains a pouch 1. As shown in Figure 11, in a state where a total of three pouches 1 are contained in the three pouch storage devices 10 that make up combination 10-1, each pouch 1 is filled with a culture medium inoculated with microorganisms to be cultured (e.g., lactic acid bacteria). In Figure 11, if the central angle of the sector in each of the three pouch storage devices 10 is set to 120°, and they are arranged adjacent to each other to form a combination 10-1 of pouch storage devices 10, then its top and bottom surfaces will be circular. However, even if the central angle θ of the sectors of the top surface 11 and bottom surface 12 is other than θ = 120°, multiple pouch storage devices 10 can be stably connected to equipment such as a culture tank 33 by arranging multiple pouch storage devices 10 side by side so that the upper and lower sectors form a circle. For example, if θ = 180°, two pouch storage devices 10 can be placed side by side so that the upper and lower sectors form a circle. Alternatively, if θ = 90°, four pouch storage devices 10 can be placed side by side so that the upper and lower sectors form a circle.

[0030] In Figure 12, a transport device 26 (a chain 26A and a gripping part 26B in the illustrated embodiment) is attached to the pouch storage device 10 to reduce the effort required to transport the pouch storage device 10 containing the pouch 1. As described above with reference to Figure 4, in Figure 12, a mounting bracket 25 is provided on the bottom surface 12 of the pouch storage device 10 (in Figure 12, the bottom surface 12 is located at the top), and one end of the chain 26A of the transport device 26 is engaged with the mounting bracket 25. The other end of the chain 26A is engaged with the gripping part 26B of the transport device 26. The engagement points of the chain 26A and the mounting bracket 25 overlap in two places in the direction perpendicular to the plane of the paper, resulting in a total of four engagement points. There are a total of two engagement points between the chain 26A and the gripping part 26B. The worker can easily transport the pouch storage device 10 containing the pouch 1 with the spout 1A facing upwards by holding the grip portion 26B of the transport device 26. In addition, the component indicated by reference numeral 22 in Figure 12 is also a handle (see Figure 6), and the handle 22 is provided for transporting the pouch storage device 10 without using the transport device 26.

[0031] Figure 13 shows a configuration in which a pouch 1 is transported by a transport device 26 (chain 26A, gripping part 26B) with a pouch connecting pipe 45 (with on / off valves 46-1 and 46-2 interposed) connected to the spout 1A of the pouch 1. In Figure 13, a fitting 25 that engages with the chain 26A of the transport device 26 is provided on the upper part of the pouch storage device 10 in Figure 13 (bottom part 12 in Figures 4 to 9), and the pouch 1 is transported by the transport device 26 with the spout 1A of the pouch 1 facing upwards in Figure 13. However, although not shown in the diagram, a fitting 25 that engages with the chain 26A is provided on the lower part of the pouch storage device 10 in Figure 13, so that the pouch 1 can be transported by the transport device 26 with the spout 1A of the pouch 1 facing downwards in Figure 13. When the pouch connection pipe 45 is connected to the spout 1A and the pouch connection pipe 45 is installed in the culture tank 33 (not shown), if the spout 1A can be grasped by the transport device 26 with the spout 1A facing downwards (downwards in Figure 13), the connection work can be performed easily and quickly.

[0032] Figure 14 shows an inoculation line 40 connecting the spout 1A of pouch 1 to the inoculation port 33A (not shown) of culture tank 33 (not shown). The inoculation line 40 has a piping system 44 which includes a connection port 41 that is selectively connected to a sterilization steam supply source 47 or a compressed air supply source 48, a tank connection port 42 that is connected to the inoculation port 33A of the culture tank 33, and an outlet 43 for sterilization steam and compressed air. The piping system 44 is connected to the pouch connection piping 45 on the pouch 1 side via a branch section 40A. The pouch connection piping 45 has two (or more) on / off valves 46-1 and 46-2 interposed therebetween, and also has a pouch connection section 45A that can be connected to the spout 1A of pouch 1. If there is a difference in diameter with the inoculation port 33A of the culture tank 33 (not shown in Figure 14), the tank connection port 42 may be structured to accommodate the difference in diameter with the inoculation port 33A. Figure 14 shows the inoculation line 40 with its various pipes (valves, connection ports, etc.) disconnected, and the inoculation line 40 is not connected to the culture tank 33.

[0033] In the illustrated embodiment, the inoculation line 40 connecting the spout 1A of pouch 1 and the inoculation port 33A of culture tank 33 is configured as shown in Figure 14. After culturing in pouch 1, the spout 1A of pouch 1 is connected to the pouch connection part 45A of the inoculation line 40, and the inoculation port 33A of culture tank 33 is connected to the tank connection port 42 of the inoculation line 40. By opening and closing the valve of the inoculation line 40, the culture medium (cultured microorganisms) in pouch 1 can be inoculated into culture tank 33 via the inoculation line 40. Compared to opening the spout 1A of pouch 1 and the inoculation port 33A of culture tank 33 and pouring the contents of pouch 1 into the inoculation port 33A, when connecting the spout 1A of pouch 1 to the inoculation line 40, the possibility of bacteria, other microorganisms, or contaminants entering pouch 1 is low, and the possibility of bacteria, other microorganisms, or contaminants entering culture tank 33 from the inoculation port 33A is also low. Therefore, the possibility of so-called "contamination" problems occurring is extremely low. Furthermore, by connecting the inoculation line 40 to the sterilization steam supply source 47 and supplying sterilization steam (high-temperature steam) and discharging it from the outlet 43, the inoculation line 40 can be sterilized. Therefore, even if bacteria enter the inoculation line 40, they will be reliably sterilized before being inoculated into the culture tank 33. Furthermore, by connecting the inoculation line 40 to the compressed air supply source 48 and discharging the supplied compressed air from the outlet 43, the inoculation line 40, which has been heated by sterilizing steam (high-temperature steam), can be cooled and its interior can be dried. Therefore, there is less risk of the microorganisms cultured in the pouch 1 being killed or damaged by the heat contained in the heated inoculation line 40.

[0034] When sterilizing the inoculation line 40, the two on-off valves 46-1 and 46-2 interposed in the pouch connection piping 45 can be closed, leaving the area between the on-off valve 46-1 on the inoculation line 40 side (culture tank 33 side) and the on-off valve 46-2 on the pouch side completely empty. Even when high-temperature steam for sterilization is passed through the inoculation line 40 in this state, the empty area between the on-off valves 46-1 and 46-2 in the pouch connection piping 45 provides insulation, preventing the high heat of the steam from being transferred to the pouch 1 side. Therefore, the culture medium (inside pouch 1) containing microorganisms cultured in pouch 1, and the culture medium located in the area near the pouch-side on / off valve 46-2 in the pouch connection piping 45, are not subjected to the high heat during sterilization. This prevents the microorganisms cultured in pouch 1 from being killed or damaged by exposure to the heat of the high-temperature steam flowing through the inoculation line 40 during inoculation line sterilization.

[0035] Pouch 1 is shown in Figure 15. In Figure 15, pouch 1 is made of a synthetic resin bag (e.g., PET / nylon / CPP) and has a predetermined heat resistance (e.g., 135°C, sterilization compatible). Microorganisms can be cultured by filling pouch 1 with culture medium. Pouch 1 is provided with a spout 1A (mouth). When inoculating into the culture tank 33, pouch 1 is placed in a pouch storage device 10 (not shown) with the spout 1A facing downwards (see Figures 1, 7, and 10). In Figure 15, the component indicated by reference numeral 27 is a cap that covers the spout 1A. Regarding the size of pouch 1, it may be set according to various standards in order to accommodate it in the pouch storage device 10 shown in Figure 4, or it can be set independently of various standards.

[0036] Next, the procedure for culturing microorganisms using the microbial culture apparatus 100, which was described with reference to Figures 1 to 15, will be explained with reference to Figure 16 and onward. In Figure 16, which shows the procedure for microbial culture in the illustrated embodiment, step S1 involves filling (injecting) the pouch 1 with a culture medium (e.g., liquid culture medium). Prior to this filling, the spout 1A of the pouch 1 is fitted into the spout attachment 23 of the pouch storage device 10 (see Figure 9), and the pouch 1 is placed in the pouch storage device 10 using the holder 24 (step of placing the pouch 1 in the pouch storage device 10). Once the process of filling pouch 1 with culture medium (e.g., liquid culture medium) is complete, the upper space (headspace) of the internal space of pouch 1 is degassed (for example, by the worker pressing on the outside of the space to degas it), and then the spout 1A is sealed tightly with the cap 27 (Figure 15). Then proceed to step S2.

[0037] In step S2, the pouch 1 filled with culture medium is sterilized (sterilization step). In the microbial culture apparatus 100 of the illustrated embodiment, a retort sterilizer (not shown) or an autoclave 32 (Figure 20) is used to sterilize the pouch 1. In the sterilization process of step S2, either a retort sterilizer or an autoclave 32 may be used. Both the retort sterilizer and the autoclave 32 can be conventionally known commercially available products (for example, the HLM-36EF model manufactured by Hirayama Seisakusho Co., Ltd. for the retort sterilizer, or the LSX-500 model manufactured by Tommy Seikou Co., Ltd. for the autoclave). In the case of a retort sterilizer, since it is equipped with a pressurizing mechanism and a cooling mechanism, even if pouch 1 is heated during sterilization, it is possible to prevent the culture medium inside pouch 1 from boiling and expanding, thus preventing pouch 1 from bursting. As an example of using a retort sterilizer, sterilization is performed at a temperature of 121°C for 30 minutes, followed by showering cooling for 30 minutes. On the other hand, since autoclaves do not have pressurizing or cooling mechanisms, heating pouch 1 in an autoclave during sterilization could cause the culture medium inside pouch 1 to boil and expand, potentially causing the pouch to burst.

[0038] Therefore, when sterilization is performed in an autoclave 32, a vent filter 35, as shown in Figure 17, is attached to the spout 1A (not shown in Figure 17) of the pouch 1 (not shown in Figure 17). When attaching the vent filter 35 of Figure 17 to the pouch 1, a vent filter mounting bracket 35-1 (made of stainless steel), as shown in Figure 18, is used. The hollow shaft 35A-1 of the vent filter mounting bracket 35-1 shown in Figure 18 is inserted into the hollow shaft 35A of the vent filter 35 shown in Figure 17, or the hollow shaft 35A of the vent filter 35 and the hollow shaft 35A-1 of the vent filter mounting bracket 35-1 are connected via a resin tube (not shown). Then, the female threaded portion (not shown) of the vent filter mounting bracket 35-1 is screwed onto the male thread on the outer circumference of the spout 1A of the pouch 1, thereby attaching the vent filter 35 to the pouch 1. The vent filter 35 ensures ventilation and maintains pressure regulation while making the sealed container or structure dustproof. By attaching the vent filter 35 to pouch 1, the pressure regulation function of the vent filter 35 is activated, preventing pouch 1 from bursting due to boiling of the culture medium. After sterilizing pouch 1, remove the vent filter 35 at the time to inoculate the culture medium inside pouch 1 with the microorganisms to be cultured.

[0039] When sterilization is performed in an autoclave 32 without a vent filter 35, a container 36 is placed inside the autoclave 32 (not shown in Figure 19) to prevent the pouch 1 from bursting during sterilization in the autoclave 32. The container 36 is open at the top and filled with water W to a predetermined level, and the pouch 1 is immersed in the container 36 filled with water W. In the inventor's experiments, as shown in Figure 19, if the water level H2 in the container 36 into which pouch 1 was immersed was 30% or more of the liquid level H1 of liquid culture medium A inside pouch 1, the rupture of pouch 1 could be prevented. In other words, the inventor's experiments have shown that the rupture of pouch 1 can be prevented if the water level H2 is 30% or more of the liquid level H1 of liquid culture medium A inside pouch 1.

[0040] Here, if the sterilization process (step S2) in Figure 16 is performed using a retort sterilizer, the use of the vent filter 35 explained in Figures 17 and 18 is unnecessary, and it is also unnecessary to immerse the pouch 1 in the container 36 containing water W as explained in Figure 19. As mentioned above, retort sterilizers are equipped with pressurizing and cooling mechanisms, which prevents the culture medium inside pouch 1 from boiling and expanding, causing pouch 1 to burst, even when pouch 1 is heated during sterilization. In the microbial culture apparatus 100 according to the illustrated embodiment, as described above, it is configured in combination with a retort sterilizer or autoclave 32 for sterilizing the pouch 1, so that the sterilization of the pouch 1 can be performed efficiently. Once pouch 1 has been sterilized, proceed to step S3.

[0041] In Figure 16, in step S3, the pouch 1, which has been filled with culture medium (e.g., liquid culture medium) (step S1) and sterilized (step S2), is inoculated with microorganisms to be cultured (e.g., lactic acid bacteria) in a clean bench, for example. Note that the pouch 1 is housed in the pouch storage device 10 at the stage of step S1, for example. Since the inoculation process in Step S3 is performed in a clean bench, the possibility of bacteria or contaminants that could cause "contamination" entering pouch 1 from spout 1A is negligibly low. Upon completion of vaccination, the upper space (headspace) inside pouch 1 is degassed, and spout 1A is sealed tightly with cap 27 (Figure 15). Next, the microorganisms inoculated into pouch 1 are cultured in an incubator (a process of culturing microorganisms in sterilized pouch 1). If the microorganisms to be cultured are lactic acid bacteria, they are cultured at a temperature of 37°C for 48 hours, for example. As for the incubator, conventionally known commercially available products (for example, model LTE-1010 from Tokyo Rikakikai Co., Ltd.) can be used.

[0042] After culturing in the incubator, proceed to step S4. In Figure 16, in step S4, the valves (on / off valves 46-1 and 46-2 shown in Figure 14) that have been sterilized in sterilization equipment (for example, an autoclave 32 or a retort sterilizer can be used) are connected to the spout 1A of pouch 1 inside a clean bench. The pouch 1 (the pouch 1 housed in the pouch storage device 10) connected to the pouch connection piping 45 is connected to the culture tank 33 via the inoculation line 40. First, connect the tank connection port 42 of the connection line 40 (the piping system 44 portion of the connection line 40) shown in Figure 14 to the inoculation port 33A of the culture tank 33. Then, sterilize the culture tank 33 and the inoculation line 40 (piping system 44). At that time, the inoculation line 40 (piping system 44) and the pouch connection pipe 45 (Figure 14) attached to the pouch 1 (spout 1A) are connected. When making the connection, as shown in Figures 1 and 2, the pouch 1 and pouch containment device 10 are installed above the culture tank 33 with the spout 1A facing downwards. Then proceed to step S5.

[0043] In the next step, S5, the inoculation line 40 connected to pouch 1 in step S4 is sterilized. For sterilization, as described above with reference to Figure 14, for example, the inoculation line 40 is selectively connected to a sterilization steam supply source 47 or a compressed air supply source 48, and sterilization steam (high-temperature steam) is supplied and discharged from the outlet 43, and compressed air is supplied and discharged. As an example, sterilization is performed by supplying high-temperature steam at 120°C for 20 minutes, followed by air cooling. When sterilizing the inoculation line 40, the two hand valves 46-1 and 46-2 interposed in the pouch connection piping 45 are closed. As a result, the area of ​​the piping between the hand valve 46-1 on the inoculation line 40 side and the hand valve 46-2 on the pouch 1 side of the pouch connection piping 45 is left empty. Even when the inoculation line 40 is sterilized at a high temperature of 120°C, the empty area of ​​the piping between the hand valves 46-1 and 46-2 in the pouch connection piping 45 prevents the high temperature of 120°C from being transferred to the pouch side. Therefore, the culture medium (inside pouch 1) containing the microorganisms cultured in pouch 1, and the culture medium located in the area near the hand valve 46-2 on the pouch 1 side of the pouch connection piping 45, are not subjected to the high heat during sterilization. As a result, the microorganisms cultured in pouch 1 are not exposed to the high-temperature steam (120°C: 20 minutes) flowing through the inoculation line 40 during inoculation line sterilization, preventing them from being killed or damaged.

[0044] After sterilizing the inoculation line 40 in step S5, proceed to step S6 and inoculate into the culture tank 33. In step S6, two hand valves 46-1 and 46-2 interposed in the pouch connection piping 45 are opened to inoculate the microorganisms cultured in pouch 1 into the culture tank 33 along with the liquid culture medium (a process of moving the microorganisms from pouch 1 into the culture tank 33). As shown in Figures 1 and 2, since pouch 1 and pouch containment device 10 are mounted above the culture tank 33 with the spout 1A on the lower side, by opening the two hand valves 46-1 and 46-2 interposed in the pouch connection piping 45, the liquid culture medium containing the microorganisms cultured in pouch 1 flows into the inoculation line 40 by gravity. Furthermore, if the viscosity of the liquid culture medium containing the cultured microorganisms is high and it is difficult for it to flow in by gravity alone, the flow can be promoted by deforming pouch 1. Once the culture medium in the pouch has flowed through the inoculation line 40 and been inoculated into the culture tank 33, close the two hand valves 46-1 and 46-2 interposed in the pouch connection pipe 45. Then, remove the pouch 1 from the inoculation line 40. However, it is also possible to disengage the pouch 1 from the pouch connection pipe 45. Next, the microorganisms inoculated from pouch 1 are cultured in culture tank 33 (step of culturing microorganisms in culture tank 33). If the microorganisms to be cultured are lactic acid bacteria, for example, the culture temperature is 37°C and the culture time is 24 hours.

[0045] The illustrated embodiments are for illustrative purposes only and are not intended to limit the technical scope of the present invention. [Explanation of Symbols]

[0046] 1. Pouch 1A... Spout 10. Pouch storage device 11...Top surface 12. Bottom 13. Radius of each of the sectors on the top and bottom surfaces 14. Each arc of a sector 15...The first straight line 16...Second straight line 32. Autoclave 33...Culture tank 33A... Inoculation port of culture tank 35...Vent filter 36...container 40... Vaccination Line 40A... Branch section 41...Connection ports for sterilization steam supply source and compressed air supply source 42... Tank connection port 43. Exhaust port for sterilization steam and compressed air. 44. Piping system connected to the pouch side via a branching point. 45. Pouch connection piping (hand valve interposed piping) 45A... Pouch connection part 46-1, 46-2... On / off valves 47. Sterilization steam supply source 48. Compressed air supply source 100...Microbial culture equipment H1... Liquid level of the culture medium in the container H2...Water level inside the container

Claims

1. A pouch containment device characterized by having a skeletal structure consisting only of the outline and outer circumference of the central part in the height direction of a hypothetical container that encloses a pouch having a spout.

2. The aforementioned skeletal structure is, The contours of the top and bottom surfaces formed in a fan shape, Two first straight lines connecting the portions of the two radii of the sectors of the upper and lower surfaces that are closer to the arc than the center, The pouch storage device according to claim 1, comprising two second straight lines connecting positions opposite to the starting point of the first straight line in each of the arcs of the sectors on the top and bottom surfaces.

3. A microbial culture apparatus characterized by having a pouch containing device according to either claim 1 or 2.

4. The pouch has an inoculation line connecting the spout and the inoculation port of the culture tank, and the inoculation line is Connection ports for the sterilization steam supply source and the compressed air supply source, The tank connection port is connected to the inoculation port of the culture tank, It has outlets for sterilizing steam and compressed air, It has a piping system that connects to the pouch side via a branching section, The microbial culture apparatus according to claim 3, wherein the piping system connected to the pouch side is connected to a pouch connection pipe, and the pouch connection pipe has a pouch connection portion that is equipped with a plurality of on / off valves and can be connected to the spout of the pouch.

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