Exchange device for microbial culture

Abstract

To provide a low-cost replacement device for microbial culture that offers good workability during replacement, reduces the workload on operators. [Solution] The replacement equipment 2 is a replacement device for microbial culture that functions as replacement equipment that can be detachably installed on the main body of the apparatus for culturing the target microorganism. It comprises a culture vessel 31 for culturing the target microorganism, a culture medium container 21 for storing consumables (culture medium B) to be replenished to the culture vessel 31 in liquid form, and a connecting part 40 and a silicone tube 33 for supplying the culture medium B from the culture medium container 21 to the culture vessel 31, all of which are pre-sterilized by gamma rays.

Description

Technical Field

[0005] , , , , ,

[0001] The present invention relates to an exchange device for microorganism culture.

Background Art

[0002] There has been a dramatic increase in the need to mass-culture microorganisms outdoors for purposes such as environmental purification technology using microorganisms such as activated sludge, microbial pesticides for protecting the diseases and health of crops, etc., adding microorganisms for preventing food poisoning bacteria in aquaculture and for the probiotic effect of seafood, and adding probiotic bacteria to the feeding of calves, piglets, etc. in the livestock field in a low-immune state for disease prevention, etc. Under such circumstances, when mass-culturing target microorganisms outdoors, it is required to culture the microorganisms in a completely sterilized container or an environment where no contamination occurs. In recent years, technologies for realizing this have been proposed (see, for example, Patent Document 1).

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0006] To achieve the above objective, an exchange device for microbial culture according to one aspect of the present invention is In a microbial culture exchange device that functions as an exchange component that is detachably installed on the main body of a device for culturing target microorganisms, A culture vessel for culturing the aforementioned target microorganism, A culture medium container for storing consumables to be replenished to the culture vessel in liquid form, A liquid delivery unit that delivers the consumables from the culture medium container to the culture vessel, The equipment is provided in a state that has been pre-sterilized by gamma rays. [Effects of the Invention]

[0007] According to the present invention, it is possible to provide a low-cost replacement device for microbial culture that offers good workability during replacement, reduces the workload on the operator, and is also cost-effective. [Brief explanation of the drawing]

[0008] [Figure 1] This figure shows an example of a service employing the on-site culture system according to the embodiment. [Figure 2] This figure shows the schematic configuration of the main unit of the on-site culture system used in the service described in Figure 1. [Figure 3] This figure shows an example of the component configuration of an on-site culture system according to the present invention. [Figure 4] Figure 3 is a side view showing a stretchable PET culture vessel applied to the on-site culture system. [Figure 5] This figure shows an example of a container receiving section applied to an on-site culture system. [Figure 6] This figure shows examples of culture medium containers made of materials that can be sterilized by gamma rays, in addition to the culture medium container shown in Figure 4. [Figure 7] This figure shows examples of culture medium containers made of materials that can be sterilized by gamma rays, in addition to the culture medium container shown in Figure 4. [Figure 8] Figure 3 is a side view showing an example of a bag-shaped culture container applied to the on-site culture system. [Figure 9] This figure shows a first example of a needle connector applied to the on-site culture system shown in Figure 3. [Figure 10] This figure shows a second example of a needle connector applied to the on-site culture system shown in Figure 3. [Figure 11] This figure shows a third example of a needle connector applied to the on-site culture system shown in Figure 3. [Figure 12] Figure 3 shows the procedure for setting the culture medium container into the cap in the on-site culture system. [Modes for carrying out the invention]

[0009] Embodiments of the present invention will be described below with reference to the drawings. First, with reference to Figure 1, we will describe a service that utilizes an on-site culture system. Figure 1 is a diagram showing an example of a service that utilizes an on-site culture system according to an embodiment.

[0010] As shown in Figure 1, this service combines the provision of an on-site culture system A to an end-user E (customer) with a business S that sells consumables regularly required for the on-site culture system A. The service provider regularly delivers refills in containers to the end-user E. The end-user E uses the refills contained in the delivered containers to culture the target microorganisms. Used containers are discarded during periodic replacements and are not returned.

[0011] The consumables used in on-site culture system A include culture media containers that hold culture media, which serve as food for microorganisms, and supplements such as drugs. These consumables are replaced regularly at a predetermined interval, for example, once a month. Furthermore, consumables include culture vessels such as large-scale culture tanks and target microorganisms such as enzyme-producing bacteria, and these are replaced periodically at the second predetermined interval, for example, once every three to four months.

[0012] In this service, the service provider gamma-sterilizes the incubator and the medium container, inoculates the target microorganism, or stores it in the incubator in the state of a pure culture inoculum, and stores the medium in the medium container, and then delivers these incubator and medium container from a predetermined delivery carrier to the end user E. The end user E sets the delivered incubator and the medium container on the outdoor microorganism culture apparatus main body, and automatically controls the culture of the target microorganism. The service provider installs the culture container and the medium container on the microorganism culture apparatus main body locally, then contracts with the end user E, and performs regular replacement by delivering the medium container storing the medium (liquid) and the culture container storing the medium (liquid) containing the microorganism (service sizing), and manages the end user E and the culture state, thereby maintaining the function as the on-site culture system A.

[0013] Thus, according to this service, by constructing a closed management system, the contamination risk (hereinafter referred to as "contamination risk") of the pure culture system can be minimized. As a result, it is possible to provide the world's first ultra-high efficiency on-site pure culture service.

[0014] In addition to the above-described service sizing-based provision method, it is also possible to provide the medium container 20 and the incubator 30 in a disposable form to be disposed of by the customer side.

[0015] Next, the configuration of the on-site culture system for realizing this service will be described with reference to FIGS. 2 and 3. FIG. 2 is a diagram showing the schematic configuration of the apparatus main body of the on-site culture system adopted in the service of FIG. 1. FIG. 3 is a diagram showing an example of the element configuration of the on-site culture system according to the embodiment.

[0016] In explaining the embodiment, "sterilization" means killing or removing all microorganisms until the survival rate is less than or equal to one in one million. "Disinfection" means killing microorganisms, but does not include the specific target microorganisms to be killed and the guarantee of the degree.

[0017] As shown in Figure 2, the on-site culture system A of this embodiment includes a control system and a liquid delivery system housing space G and a culture space F inside the housing D. Culture space F houses culture vessels 20 and 30 in a removable manner, as well as capsule filters, ventilation fans, temperature sensors, and other components. The culture medium container 20 is set in the container holder 41 fixed to the housing D. The incubator 30 is placed on a rubber heater (not shown) at the bottom. The temperature of the culture medium inside the incubator 30 is controlled by the rubber heater. Due to its weight, enclosure D is equipped with lifting rings (locking parts) at the four corners of its top for lifting with a crane or similar equipment. Casters for fine-tuning the position of enclosure D, as well as adjustable fixing devices for adjusting and securing the enclosure D's level, are located at the four corners of its bottom.

[0018] The control system and fluid supply system contain space G, which houses the control unit 10 shown in Figure 3 and the equipment to be controlled. The equipment to be controlled includes, for example, compressors, temperature control controllers, water supply solenoid valves, flow meters, tubing pumps, air pumps and other pumps, and tubing for fluid and air supply.

[0019] As shown in Figure 3, the on-site culture system A of this embodiment comprises a main unit 1 and replacement equipment 2.

[0020] The main unit 1 of the apparatus is housed in an outdoor-type enclosure and includes a blower 11, pressure regulators 12a and 12b, flow meters 13a and 13b, solenoid valve 14, tube connectors 15a to 15i, a membrane filter 16 for air sterilization, a capsule filter 39 for water filtration sterilization, a container receiving section 41, tubing pumps 17 and 18 for liquid transfer, a control unit 10, a rubber heater (not shown), a cooling device such as a cooler (not shown), and the like.

[0021] The control unit 10 controls the equipment to be controlled (blower 11, pressure regulators 12a, 12b, tubing pumps 17, 18, etc.). The control unit 10 includes a touch panel 10a, a PLC (Programmable Logic Controller) 10b, a power breaker 10c, and the like. The touch panel 10a is operated by an operator (worker) via touch, allowing for various settings and instructions to be given to the controlled equipment. The PLC10b incorporates a microprocessor and controls the target devices through user-modifiable programs. Furthermore, the PLC10b controls the target devices according to various settings and touch operations. The power breaker-10c shuts off the power supply to the controller 10 and the controlled equipment when an overcurrent is detected.

[0022] The blower 11 is the source (supply source) of aeration air and is controlled by the PLC 10b of the control unit 10 to supply aeration air to the culture vessel 30 on the exchange equipment 2 side. The rotational speed of the tubing pumps 17 and 18 is controlled by the PLC 10b of the control unit 10, thereby adjusting the fluid flow rate. In addition, the fluid flow rate of the tubing pump 18 is adjusted by the inner diameter of the tube and the operating time.

[0023] The pressure regulator 12a adjusts the pressure (flow rate) of the air used for aeration. The flow meter 13a measures the air flow rate and inputs the measured flow rate to the controller 10. The pressure regulator 12b adjusts the pressure (flow rate) of the city water injected from the outside through the tube connector 15g of the main unit 1. The flow meter 13b measures the flow rate of the city water, etc., and inputs the measured flow rate to the controller 10. City water refers to tap water, but other types of water can also be used, so in this explanation, it is referred to as city water, etc. The solenoid valve 14 is a valve for supplying and stopping the supply of tap water to the culture vessel 30. The solenoid valve 14 is controlled by the controller 10 (PLC10b, etc.) to be ON / OFF, etc. The rubber heater is positioned at the bottom of the culture container 31 of the incubator 30 on the replacement equipment 2 side and is used to raise the temperature of the incubator 30. The rubber heater has a temperature control function that includes a temperature sensor, and is capable of maintaining the temperature of the liquid in the culture container 31 within a certain temperature range. Furthermore, if cooling is required, the temperature can be lowered by passing a cooling pipe containing a refrigerant such as cold water inside or outside the culture vessel 31. Additionally, the temperature can be lowered by attaching a cooler to the enclosure. By using both the rubber heater and the cooler in combination, the temperature of the on-site culture system A can be controlled to a constant level. The membrane filter 16 for air sterilization sterilizes the air taken into the main body 1 of the device. The capsule filter 39 for water filtration sterilization sterilizes tap water or other public water taken into the main body 1 of the device.

[0024] The replacement equipment 2 includes, for example, a culture medium 20 weighing 130g and with a capacity of 12L (liters), and an incubator 30 with a capacity of, for example, 20L (liters). All of these containers are made of materials that can be sterilized by gamma rays. Specifically, materials such as nylon, polyethylene, polyethylene terephthalate, polycarbonate, and polypropylene are used. As a result, both the culture medium 20 and the incubator 30 are very lightweight, reducing the workload on the user and making disposal after use easier.

[0025] The culture medium container 20 has a container body, such as a culture medium container 21, in which the culture medium (liquid) B is stored. The culture medium (liquid) B is a liquid containing components (culture medium components) that serve as food for the target microorganisms. As shown in Figure 4, the culture medium container 21 is a gamma-ray sterilized stretchable polyethylene terephthalate container (stretchable PET container). The stretchable PET container has the external shape of a gallon bottle, such as those used in water dispensers, and is fixed to the main body of the device 1 by inverting it and inserting the lower end (at least the conical part including the cap 23) into the container receiving part 41.

[0026] Now, with reference to Figure 5, the container receiving portion used in the connection part of Figure 3 will be described. Figure 5 shows an example of a container receiving part used in the connection part shown in Figure 3.

[0027] As shown in Figure 5, the container receiving portion 41, which constitutes part of the connecting portion 40 (see Figure 3), has a recess 41a that is opened in a substantially annular shape, a cup portion 41b (medication supply portion) that protrudes radially from a part of the peripheral edge of the recess 41a, a spill 41c, a bamboo shoot 41d for connecting a silicone tube, and a silicone cap 41e. In other words, the container receiving section 41 has an opening that has the shape of a keyhole-shaped burial mound, and its inner wall is shaped to receive the lower end portion (at least the conical portion including the cap 23) of the inverted gallon bottle-shaped culture medium container 21. In the case of the container receiving section 41, the recess 41a has an opening at the top. With the cap 23 screwed onto the threaded opening 21a (see Figure 4) at the end of the culture medium container 21, the cap 23 is inserted downwards into the recess 41a. This causes the spill 41c protruding from the bottom of the recess 41a to penetrate the rubber stopper portion of the cap 23, thereby connecting the culture medium container 21 and the container receiving section 41. In the case of the container receiving section 41, with the culture medium container 21 placed in the container receiving section 41, disinfectants such as sterilizing liquid or sterilizing gel can be added (replenished) through the opening of the cup section 41b without lifting the culture medium container 21.

[0028] The spill 41c is positioned in the center of the recess 41a, and with the culture medium container 21 in place, the culture medium in the culture medium container 21 can be supplied to the culture vessel 31 below through the silicone tube 33 (see Figure 3) via a hole provided at its tip, through which the culture medium in the culture medium container 21 is routed to the bamboo shoot 41d for connecting the silicone tube. The silicone cap 41e (lid) is fitted to cover the periphery of the opening of the recess 41a, sealing the opening, and its appearance is shaped like a keyhole-shaped burial mound, similar to the opening of the recess 41a. This prevents bacteria and other contaminants from entering the recess 41a, maintaining a sterile state inside the recess 41a.

[0029] By making the shape of the opening of the recess 41a of the container receiving portion 41 a keyhole-shaped mound, sterilizing liquid or sterilizing gel can be added (replenished) from the protruding cup portion 41b, and the recess 41a can always be filled with disinfectant. Therefore, the spill 41c located inside the recess 41a can be kept in a sterilized state at all times.

[0030] In particular, when changing the culture medium, performing the change while the container is filled with sterilization solution or gel allows for easy connection to the culture medium container 21 while maintaining a sterile condition.

[0031] In this example, a silicone tube 33 for connecting to the culture vessel 31 is connected to the lower part of the spill 41c of the connection part 40 to the culture medium container 21. Preferably, a connector with a stopcock is attached to the silicone tube 33 to make it easier to handle sterilization solutions and sterilization gels.

[0032] A cap 23 with a rubber stopper is screwed onto the threaded opening 21a (also called the threaded portion or opening) at the lower end of the culture medium container 21. The cap 23 with the rubber stopper is part of the connection portion 40. In this way, by using a stretchable PET container for the culture medium container 21, negative pressure is created inside the stretchable PET as the culture medium is supplied, causing the container itself to collapse naturally. This makes it possible to transport the culture medium using a lighter material and dramatically improves maintainability by making it less bulky when disposed of after use. The culture medium container 21 is pre-filled with culture medium components suitable for the microorganism to be cultured. It is preferable to fill it with a concentrated culture medium solution. This is because it extends the replacement interval and, by concentrating the culture medium components, lowers the water activity value, thereby reducing the risk of freezing point depression and contamination by unwanted bacteria. The culture medium (liquid) B in the culture medium container 21 is transferred to the culture vessel 31 through the silicone tube 33 and the connecting part 40 connected to the culture vessel 31.

[0033] In addition to the culture medium container 21 made of the stretchable PET described above, other containers made of materials that can be sterilized by gamma rays, such as intravenous fluid bags for drip infusions, can also be used. Specifically, as shown in Figure 6, for example, single-use bags 51 to 55 made of polyethylene (PE) or nylon material, or cubitainers 61 to 63 (bag-in-box) as shown in Figure 7, can also be used as culture medium containers 21, and their capacity and shape can be freely designed.

[0034] Now, let's describe the culture vessel with reference to Figure 8. Figure 8 shows an example of a culture vessel. The culture vessel 30 described above has a container body made of PE with a side shape as shown in Figure 8, such as a culture vessel 31 (for example, the shape of an infusion pack such as an intravenous drip pack). The culture vessel 31 includes a hanging section 71 and a culture medium storage section 72. The hanging section 71 is provided with a hanging hook hole 73. The culture medium storage section 72 is provided with a sterile water / culture medium inlet 74, a culture medium inlet 75, an overflow outlet 76, an aeration port 77, a culture medium outlet 78, etc. PE tube connectors are attached to the openings other than the hanging hook hole 73.

[0035] By using a PE bag-shaped culture container 31 in this way, inoculation after gamma ray sterilization becomes easier, and the bulk and weight of transportation to the site are greatly reduced, thus lowering transportation costs and reducing the labor burden on users for installation.

[0036] While typical culture containers made of SUS304 or PP require a minimum volume of 20L, the PE culture container 31 used in this embodiment can be transported simply by adding 100mL of starter culture to the bag, thus significantly reducing transport volume and weight.

[0037] At the destination, liquid C (hereinafter abbreviated as "culture solution C") containing culture medium (liquid) B from the culture medium container 20 set in the main unit 1 and 100 mL of the target microorganism that has been inoculated in advance is placed in the culture container 31.

[0038] The culture vessel 31 is fitted with piping such as silicone tubes or Teflon tubes for liquid or air passage. Note that Teflon is a registered trademark. The culture vessel 31 is fitted with, for example, a silicone tube 38 for supplying aeration air and a silicone tube 33 for passing the culture medium.

[0039] A rubber heater is provided at the bottom of the replacement substrate 2. The rubber heater has a temperature control function, such as a temperature-controlled rubber heater, and is located at the bottom of the culture vessel 31. The temperature-controlled rubber heater can adjust the culture temperature to the optimal temperature suitable for each microorganism being cultured. Generally, the growth rate of microorganisms is maximized in the range of 20°C to 38°C, and heaters that allow fine temperature adjustment within this range are used. For example, the culture temperature for the lipid-degrading bacterium Rhodococcus erythropolis is about 20°C, while Pseudomonas putida shows high activity at about 35°C.

[0040] A tube connector 15a is provided at one end of the water supply tube 36. A tube connector 15b is provided at one end of the aeration air supply tube 38. The culture medium outlet tube 37 is a tube for discharging the liquid containing microorganisms (target microorganisms) as culture medium C to the outside of the culture vessel 31, and a tube connector 15c is provided at one end of the culture medium outlet tube 37. A tube connector 15d is provided at one end of the overflow tube 35. These tube connectors 15a to 15d are designed to be easily attached to and detached from the main body of the device 1 and the piping (tube components, etc.) of external equipment.

[0041] Air is supplied to the culture vessel 31 through an aeration air supply tube 38. An air sterilization membrane filter 16 (capsule filter) is attached to the aeration air supply tube 38, and the air is sterilized by filtration.

[0042] The blower 11 installed in the main body 1 of the apparatus is used as the source (supply) of aeration air. Alternatively, an air pump or the like may be used. Between the blower 11 and the culture vessel 31, a pressure regulator 12a, a flow meter 13a, a membrane filter 16 for air sterilization with a pore size of, for example, 0.2 μm, an aeration air supply tube 38, and a tube connector 15b are connected in that order from the blower 11 side, forming an air flow path. The components from blower 11 to tube connector 15i are the equipment on the main unit 1 side of the device.

[0043] The aeration air introduced into the main body 1 of the device by the blower 11 is introduced into the culture container 31 through the air passage via a removable tube connector 15b attached to the culture container 31. Direct aeration alone is sufficient, but a diffuser can also be attached to the bottom of the culture container 31 to aerate the culture medium C. Diluting the air with a diffuser or the like improves the oxygen dissolution efficiency, thereby improving the culture efficiency.

[0044] The culture vessel 31 is supplied with tap water (city water) for diluting the culture medium via a tube connector 15a through a city water supply tube 36. The city water supply tube 36 is a silicone tube. Note that any liquid that can be filtered and sterilized and does not adversely affect the culture of microorganisms, such as well water or industrial water, is acceptable instead of tap water. Therefore, the water is filtered and sterilized using a capsule filter 39 with a pore size of 0.45 μm (preferably 0.2 μm or less). In this example, a water sterilization capsule filter with a pore size of 0.2 μm is used as the capsule filter 39. Other methods, although at a sterilization level, may include sterilization techniques such as ultraviolet sterilization, heat sterilization, high frequency, microwave, ultrasound, ozone, or plasma.

[0045] The device is equipped with a pressure regulator 12b, a flow meter 13b, a solenoid valve 14, a capsule filter 39, and a water supply tube 36 as means of supplying tap water. Tap water introduced into the main body 1 is supplied to the culture container 31 through the above supply means via a removable tube connector 15a connected to one end of the water supply tube 36. In order to filter and sterilize the tap water with the capsule filter 39, a water pressure of approximately 0.3 MPa or higher is required on the raw water side. If the water pressure is low, it shall be pressurized separately with a pump or the like. The tap water is introduced into the culture container 31 through the above water supply route.

[0046] The culture medium C cultured in the culture vessel 31 is transferred from the tube connector 15c connected to the culture medium outlet 78 (see Figure 8) through the culture medium outlet tube 37 to the outside of the apparatus via the tube connector 15e, and added to the required locations. The culture medium outlet 78 is positioned slightly above the bottom of the culture vessel 31 so that approximately 1-5% of the total volume of culture medium C remains in the culture vessel 31.

[0047] The culture medium outlet tube 37 attached to the culture vessel 31 is a pipe for transferring the culture medium C inside the culture vessel 31 to the outside of the device. The culture medium C is drawn out via the culture medium outlet tube 37 by a tubing pump 18 installed in the main body of the device 1 and transferred to an external destination (addition destination). The rotation speed of the tubing pumps 17 and 18 is controlled by the PLC 10b, which adjusts the flow rate of culture medium B and culture solution C. In addition, the flow rate can also be adjusted by the inner diameter of the tube and the operating time set by the program timer.

[0048] The tube connector 15d attached to the overflow outlet 76 (see Figure 8) of the culture vessel 31 is connected to the tube connector 15f via the overflow tube 35, and any excess overflow is discharged from the tube connector 15f. The overflow tube 35 is laid in a location where it is not a problem if a small amount of cultured microorganisms and culture medium components flow out, such as where the culture medium (liquid) is added or in a sewage pipe line. Furthermore, to prevent contamination due to backflow (hereinafter referred to as "contamination"), it is desirable to connect it to a container filled with a small amount of sterilizing / disinfecting agent and leave it open under aeration.

[0049] The overflow tube 35 is installed to prevent the internal pressure inside the culture vessel 31 from rising. Specifically, the overflow tube 35 functions as an outlet for aeration air used for culturing, a channel for bubbles to escape if foaming occurs during culturing, and a channel for the culture medium C to escape if too much tap water or culture medium is added due to some abnormality.

[0050] The culture vessel 31 and culture medium container 21 set described above are sterilized by gamma rays, then sealed in a vacuum pack or the like, and transported to the site where the main unit 1 is installed.

[0051] The connection of replacement equipment 2, particularly the tubes between the culture vessel 31 and the main unit 1 of the device, can be replaced with alcohol sterilization only, depending on the risk of contamination. When replacing replacement equipment 2, the tube connectors 15h, 15i, 15e, and 15f are inserted directly into each port of the culture vessel 31.

[0052] The operation of the main unit 1 is controlled by a controller 10 (PLC10b, etc.) installed on the main unit 1. The controller primarily controls the ON / OFF state of pumps and solenoid valves, and the operation sequence of timers, etc., thereby controlling the culture / addition process, the culture medium exchange process, and the incubator exchange process. Furthermore, by equipping the PLC10b with a communication line function, it becomes possible to remotely control the device from an external source, check the device status, and verify the occurrence of errors.

[0053] In addition, the incubator 30, including the culture vessel 31, may be equipped with stirring components (stirring blades, stirring rotors, etc.), a pH meter, a DO meter, an alkali injection pump, an alkali inlet, an acid injection pump, an acid inlet, an electrode-type water level sensor, a turbidity sensor, a cooler, and the like.

[0054] A connector 40 is provided at one end of the silicone tube 33 for passing the culture medium through, for sterile connection to the culture medium container 21.

[0055] Here, another example of the connection part 40 will be described with reference to Figure 9. Figure 9 shows a connection structure different from the connection part 40 shown in Figure 3 (a case without the container receiving part 41). In this structure, a cap 23 with a rubber stopper is screwed onto the threaded opening 21a at the lower end of the culture medium container 21 shown in Figure 4, and the needle connector 42 is directly inserted into the rubber stopper portion of the cap 23 to make the connection. In this case, the culture medium container 21 is suspended from the main body of the device 1 in an inverted state.

[0056] Specifically, as shown in Figure 9, the needle connector 42 comprises a base portion 43, a needle portion 44, a cup portion 45 (disinfectant storage portion), and a cap 46. The dashed line shown in the figure indicates the liquid level of the disinfectant 48. The base portion 43 is a base that supports the needle portion 44, and a bamboo shoot 47 (cylindrical portion) for connecting the silicone tube 33 is provided below it. The needle portion 44 is tubular with a pointed tip, resembling an injection needle, and penetrates the base portion 43. The needle portion 44 is connected to the culture medium container 21 by inserting the tip of the injection needle-shaped portion into the cap 23 on the culture medium container 21 side. The needle portion 44 is provided at one end of the silicone tube 33 from the culture vessel 31. The needle portion 44 is inserted into the rubber stoppered cap 23 at the lower end of the bottle-type culture medium container 21, which is supported by the container receiving portion 41 (see Figures 3 and 5). The cup portion 45 houses the needle connector 42 and, for example, is made of a transparent resin so that the needle tip can be visually inspected. Alternatively, the material of the cup portion 45 may be metal, solid resin, flexible resin, etc. In other words, the cup portion 45 can be made of any material as long as it is a container-like structure into which the connecting portion can be immersed.

[0057] The cup portion 45 is positioned (in a cylindrical shape) along the edge of the base portion 43 so as to surround the needle portion 44, and covers up to a height higher than the needle portion 44. The cup portion 45 and the base portion 43 together form a cylindrical container. The cup portion 45 contains a disinfectant 48 into which the needle connector 42 is immersed. Specifically, a liquid or gel-like disinfectant 48 is stored (filled) in a cylindrical container that makes up the cup portion 45. Examples of disinfectants 48 that can be used include 70% ethanol, peracetic acid, glutaraldehyde, etc.

[0058] The cap 46 is a lid designed to prevent the disinfectant 48 inside the cup 45 from spilling out during transport or installation.

[0059] In a connection structure like this example, the opening of the cup portion 45 is sealed by the cap 46 during transport, so the disinfectant 48 contained in the cup portion 45 does not spill out during transport or installation. Furthermore, when setting up the culture medium container 21, the cap 46 is removed and the tip of the needle portion 44 is inserted into the cap 23 at the lower end of the culture medium container 21. At this point, the cap 23 and the tip of the needle portion 44 are immersed in the disinfectant 48, and the cap 23 and the needle portion 44 are connected in a sterile state, thus preventing the entry of bacteria from the connection portion 40. In other words, bacterial contamination during container replacement can be prevented.

[0060] In the above embodiment, the structure in which the spill 41c of the container receiving portion 41 and the needle connector 42 are inserted into the rubber stoppered cap 23 on the culture medium container 21 side during connection was described as an example of the connection portion 40, but other configurations are also possible. For example, as shown in Figure 10, a bag-shaped infusion pack made of PE or nylon material, used for medical purposes, may be used as the culture medium container 21, and the needle portion 44 of the needle connector 42 may be inserted into the rubber stoppered connector 91 at its lower end. The needle connector 42 shown here is an example in which the base portion 43, the needle portion 44, and the cup portion 45 are integrally molded from, for example, resin.

[0061] Alternatively, as shown in Figure 11, the injection port 101 with a sterilized rubber stopper may be inserted into the needle connector 42.

[0062] In this way, by connecting the needle connector 42 in a sterile or sterilizing solution, a sterilized state is always maintained, allowing the culture medium to be replaced repeatedly.

[0063] Next, with reference to Figure 12, a first modified example of the cap used for the connection part in Figure 3 will be described. Figure 12 shows a modified example of the cap used for the connection part in Figure 3. The cap used for the connection in Figure 3 was a cap with a rubber stopper, but other types of caps may also be used. For example, as shown in Figure 12, the modified cap 23 comprises a threaded portion (not shown) that is screwed onto a threaded opening 21a (liquid outlet) for dispensing the culture medium B (liquid) contained in the culture medium container 21, a recess 23a formed on the outside of the threaded portion (on the cap surface side), absorbent cotton 112 that is contained in the recess 23a and moistened (impregnated) with a sterilizing solution, and a top lid seal 111 (sealing member) that seals the opening of the recess 23a with the absorbent cotton 112 contained in the recess 23a.

[0064] In the case of a cap 23 with this configuration, in step S11 of Figure 12, after filling the culture medium (liquid) B into the stretchable PET culture medium container 21, cotton wool 112 moistened with sterilizing alcohol (alcohol concentration 60% to 90%, preferably 70% to 85%) is placed in the recess 23a of the cap 23, and the top lid seal 111 is attached to close the opening of the recess 23a to seal the recess 23a. After that, gamma ray sterilization is performed. The culture medium container 20, modified in this manner, is used as a replacement culture medium. Therefore, the filter for air supply that was previously required is no longer necessary.

[0065] During the culture medium change in step S12, even when the top lid seal 111 is peeled back, the recessed area 23a remains moist with the alcohol-soaked cotton wool 112, and the sterilization effect is maintained. Therefore, during the time the culture medium is changed in step S13 (for example, 1 to 5 minutes), the recess 23a of the cap 23 remains sterile.

[0066] This eliminates the need to provide a special drive mechanism for connector connection in the connection section 40, thereby simplifying the device and reducing costs, while also eliminating the technical burden on replacement workers and dramatically improving maintainability.

[0067] Furthermore, the disinfectant used to moisten the cotton wool 112 contained in the recess 23a of the cap 23 may be a liquid disinfectant with bactericidal or sterilizing properties other than alcohol, such as peracetic acid, glutaraldehyde, or iodine. In addition, a gel-type disinfectant or sterilizing solution can be used as a substitute for the cotton wool 112.

[0068] Here, we report the results of an experiment conducted to confirm that no contamination occurs when changing the culture medium container 21, using a cap 23 containing absorbent cotton 112 moistened with sterilizing alcohol (70%).

[0069] The experiment was carried out according to the following steps 1 through 3. 1. Peel back the top lid seal 111 of the cap 23 of the culture medium container 21 and moisten it with a 70% alcohol swab. 2. Afterwards, it was left outdoors for 1 minute. 3. After being left outdoors, the samples were transferred to a clean bench, approximately 3 mL of SCD medium was poured into the caps, and the samples were incubated at room temperature for 4 days to check whether colonies were formed on the inside of the caps (connection ports). The above test was performed on 10 caps (N=10), and no bacterial growth was observed inside any of the caps. Based on these experimental results, it was concluded that by filling the inside of the cap 23 with a disinfectant solution or disinfectant gel, the disinfectant effect is maintained and contamination does not occur, even when the connection between the culture medium container 21 and the culture vessel 31 is temporarily opened during culture medium replacement, as long as the moisture state of the cap 23 is maintained.

[0070] As described above, according to this embodiment, by forming the culture medium container 21 and the culture vessel 31 from a lightweight and highly elastic material, such as nylon, polyethylene, polyethylene terephthalate, polycarbonate, or polypropylene, both the culture medium container 21 and the culture vessel 31 become extremely lightweight, reducing the workload on the user and making disposal after use easier. Furthermore, by making the connection part 40 that connects the culture medium container 21 and the culture vessel 31 a one-touch structure (inserting the needle connector 42 into the cap 23), the culture medium container 21 can be replaced without giving any opportunity for bacteria or other contaminants to enter. In other words, contamination of the culture medium during container replacement can be prevented. Furthermore, by using a stretchable PET bottle for the culture medium container 21, negative pressure is created inside the stretchable PET as the culture medium is supplied, causing it to collapse naturally. This allows for the transport of the culture medium using a lighter material, and significantly improves maintainability by reducing the bulk during disposal after use.

[0071] The following describes the installation process and operation of the exchange equipment (incubator and culture medium) in the on-site culture system of the embodiment, referring to the diagram described above.

[0072] At the supplier of replacement equipment 2, the membrane filter 16 for air sterilization and the water supply tube 36 connected to the capsule filter 39 for water sterilization are connected to the culture vessel 31. In Figure 8, a tube connector 15d, to which an overflow tube 35 is connected, is connected to the overflow outlet 76 of the culture vessel 31. A tube connector 15a, to which a silicone tube is connected, and a tube connector 15c, to which a culture outlet tube 37 (Pharmamed tube for the tubing pump) is connected, are also connected to the sterile water / culture medium inlet 74 and the culture solution outlet 78, respectively.

[0073] In this state, all instruments are sterilized by gamma rays to complete the process. Then, before shipment, 100-1000 mL of the target culture microorganism is filled into culture vessel 31, and the vessel is delivered to the end user E's site of use.

[0074] On-site, the culture vessel 31 is placed in the housing D of the main unit 1 (see Figure 2). Specifically, the culture vessel 31 is placed in the culture space F of housing D. Then, the circuit breaker of the main unit 1 is turned ON. At this time, only the compressor 19 of the main unit 1 is started.

[0075] Next, the membrane filter 16 for air sterilization is connected to a tube (not shown) that is connected to the compressor 19 in Figure 2. A capsule filter 39 for water sterilization is connected to the tube at the outlet of the solenoid valve 14 for water supply. The overflow tube 35 is connected to the silicone tube (not shown) at the bottom of the housing D to which the tube connector 15f is connected. The culture medium outlet tube 37, which is connected to the culture medium outlet 78 of the culture vessel 31 in Figure 8, is set on the tubing pump 18 and connected to the transfer port tube connector 15e. To prevent the culture medium or disinfectant solution from flowing downstream, either clip the silicone tube 33 with a clip or similar component, or, if the silicone tube 33 has a connector with a stopcock, set the container receiving section 41 with the stopcock closed onto 40. At this point, the container receiving section 41 is already filled with disinfectant solution or gel. Set the silicone tube of the culture medium inlet 74 of the culture vessel 31 in Figure 8 onto the tubing pump 17. Then, remove the clip from the silicone tube 33 or open the stopcock. Here, the culture vessel 31 set in the housing D can be used for several months or more, depending on the lifespan of the capsule filter 39 for air sterilization.

[0076] Then, fill the container receiving section 41 with sterilizing alcohol (60-90%, preferably 70-85%), and insert the inverted culture medium container 21 into the container receiving section 41 as shown in Figure 4, in the same manner as setting up a water dispenser, and fit the cap 23 onto the spill 41c of the container receiving section 41. In this case, if there is insufficient disinfectant alcohol filled in the recess 41a of the container receiving section 41, disinfectant alcohol is replenished from the cup section 41b of the container receiving section 41 to fill the recess 41a with disinfectant alcohol. In addition to alcohol, other liquid agents with disinfectant properties such as peracetic acid, glutaraldehyde, and iodine, or disinfectant solutions in gel form may be used as substitutes. The culture medium container 21 is replaced once to several times a month, depending on the container's capacity and the amount used. At that time, the container receiving section 41 is filled with sterilizing alcohol.

[0077] If the connection part 40 is to be used with a combination of, for example, the cap 23 in Figure 12 and the needle connector 42 in Figure 9, instead of the combination of the cap 23 and the container receiving part 41 in Figure 3, then the cap 23 in Figure 12 is screwed onto the threaded opening 21a of the culture medium container 21, as shown in Figure 4. In this case, after peeling off the top lid seal 111 of the cap 23 in the procedure shown in Figure 12, the needle part 44 of the needle connector 42 is inserted into the recess 23a of the cap 23.

[0078] As described above, after setting the replacement substrate 2 in the main unit 1 of the apparatus, the operator presses the culture start button on the touch panel 10a of the control unit 10 in Figure 3, and sterile water and culture medium are dispensed in the set arbitrary volume. Sterilized water is supplied to the main unit 1 of the apparatus at a pressure of 0.2 to 0.6 MPa, the amount of water is measured by the flow meter 13b in Figure 3, and the water supply solenoid valve 14 is opened and closed.

[0079] The amount of culture medium B added can be adjusted by controlling the operating time of the tubing pump 17. The air flow rate from the compressor 19 (see Figure 2) can be arbitrarily adjusted by the flow meter 13a. Subsequently, incubation for the set time with aeration begins.

[0080] After the culturing is complete, the tubing pump 18 shown in Figure 3 operates for a pre-set time to transfer (add) the culture medium C in the culture vessel 31 to an external location (any location). If the capacity of the culture medium container 21 is smaller than the capacity of the culture vessel 31, the exchange of the cap 23 and the culture medium container 21 will be repeated until the culture vessel 31 is replaced. When the culture vessel 31 is replaced, the above procedure and operations for the culture vessel 31 are repeated.

[0081] As described above, according to this embodiment, by using expandable PET bottles for the culture vessel 31 and culture medium container 21, and transporting them after gamma ray sterilization for delivery to the end user E, both the culture medium container 21 and culture vessel 31 become very lightweight, reducing transportation costs, decreasing the workload on the user, and making disposal after use easier.

[0082] The embodiments described above are merely examples of how to carry out the present invention, and the present invention is not limited to the embodiments described above.

[0083] In the above embodiment, stretchable PET bottles (see Figure 4) were used as containers for the culture vessel 31 and the culture medium container 21, but other containers made of stretchable and gamma-ray sterilizable material may also be used.

[0084] In other words, the exchange device for microbial culture to which the present invention is applied only needs to have the following configuration, and can take various forms. (1) That is, the exchange device for microbial culture to which the present invention is applied (for example, exchange device 2 in Figure 3, etc.) In a microbial culture exchange device that functions as an exchange device (e.g., exchange device 2 in Figure 3) that is detachably installed on the main body of the apparatus for culturing target microorganisms (e.g., main body 1 in Figure 3), A culture vessel for culturing the target microorganism (for example, the microorganism in culture medium C, etc.) (for example, culture vessel 31 in Figures 3 and 8, etc.), A culture medium container (for example, culture medium container 21 in Figures 3 and 4, single-use bags 51 to 55 in Figure 6, or cubitainers 61 to 63 in Figure 7, etc.) that stores consumables (for example, culture medium B, chemicals, etc.) to be replenished to the culture vessel in liquid form, A liquid delivery unit (for example, the silicone tube 33 for liquid delivery of the culture medium shown in Figure 3) delivers the consumables (culture medium B, chemicals, etc.) from the culture medium container to the culture vessel, The equipment is provided in a state that has been pre-sterilized by gamma rays. In this way, by providing the culture container (e.g., culture container 31 in Figures 3 and 8), the culture medium container (e.g., culture medium container 21 in Figure 3), and the liquid delivery unit (e.g., silicone tube 33 for culture medium delivery in Figure 3) of the replacement equipment (e.g., replacement equipment 2 in Figure 3) that is detachably installed on the main body of the apparatus for culturing the target microorganism (e.g., main body 1 in Figure 3), in a state where they have been pre-sterilized with gamma rays, it is possible to provide containers made of lightweight and highly elastic materials, such as nylon, polyethylene, polyethylene terephthalate, polycarbonate, and polypropylene. This makes both the culture medium container and the culture vessel extremely lightweight, reducing the workload for the user and making post-use disposal (e.g., waste disposal) easier. Furthermore, the entire exchange system can be designed to prevent contamination by preventing the growth and entry of microorganisms other than the target microorganism.

[0085] (2) In the above-mentioned exchange device (for example, exchange equipment 2 in Figure 3), The culture vessel has a bag-like shape (for example, the shape of a PE drip bag). This makes inoculation after gamma ray sterilization easier and significantly reduces the bulk and weight of transport to the site, thereby lowering transportation costs and reducing the labor burden on users for installation. In other words, the culture containers that hold the culture medium can be made more compact, reducing transportation costs. Also, since used culture containers can be disposed of when they are replaced, transportation costs for container collection (return) can also be reduced.

[0086] (3) In the above-mentioned exchange device (for example, exchange equipment 2 in Figure 3), The culture vessel has tube connectors (e.g., tube connectors 15a to 15d in Figure 3, etc.) for connecting other tubes to the infusion pack (e.g., culture vessel 31 in Figure 8). This allows the tube connector to be connected to other tubes or culture medium containers 21 in a sterile state, thus preventing contamination and saving space.

[0087] (4) The culture medium container (for example, culture medium container 21 in Figures 3 and 4) is made of a shrinkable container (polyethylene terephthalate: PET container, etc.). In this way, by using a shrinkable container (such as a PET bottle) for the culture medium, negative pressure is created inside the expandable PET as the culture medium is supplied, causing the container to collapse naturally. This allows for the transport of the culture medium using lighter materials, and significantly improves maintainability by reducing the bulk of the container during disposal after use.

[0088] (5) The culture medium container is The container body (for example, culture medium container 21 in Figures 3 and 4), A liquid outlet (screw opening 21a in Figure 4) for dispensing the culture medium (e.g., culture medium B, etc.) contained within the container body, A cap (for example, the cap 23 in Figures 3 and 4) having a threaded portion that is screwed into the liquid outlet and a recess formed on the back side of that portion (recess 23a in Figure 12), The cap contains a cotton ball moistened with a disinfectant solution (for example, cotton ball 112 in Figure 12), and the cap contains a cotton ball moistened with a disinfectant solution. It is equipped with. This eliminates the need for the air supply filter that was previously required. Furthermore, even when the top lid seal is peeled back during culture medium replacement, the alcohol swab remains moist, maintaining its sterilizing effect. This confirms that the sterilization system can be maintained even during culture medium replacement (for example, for about 1 to 5 minutes) (based on experimental results). As a result, a special drive mechanism is no longer required in the part that receives the culture medium container 21, achieving simplification and cost reduction of the device, while also dramatically improving the technical burden on the user and ease of maintenance. In addition to alcohol swabs, the disinfectant to be filled into the cap opening can also be a liquid disinfectant with bactericidal or sterilizing properties such as peracetic acid, glutaraldehyde, or iodine, or a gel-type disinfectant or sterilizing solution can be used instead of swabs.

[0089] (6) The system is equipped with a support part (for example, the container receiving part 41 in Figure 5) that supports the culture medium container (for example, the culture medium container 21 in Figures 3 and 4), The aforementioned support portion (for example, the container receiving portion 41 in Figure 5) A sterilizing solution (e.g., a disinfectant) is filled into a recess (e.g., recess 41a in Figure 5) in which the culture medium extraction part (e.g., cap 23 in Figures 3 and 4) of the culture medium container (e.g., culture medium container 21 in Figures 3 and 4) is housed so as to be immersed in the sterilizing solution, The disinfectant liquid supply section (for example, the cup section 41b in Figure 5) is provided to protrude radially from the edge surrounding the recess (for example, the recess 41a in Figure 5), It is equipped with. This allows for the addition (replenishment) of sterilizing / disinfecting solution or gel from the radially protruding supply section (e.g., cup section 41b in Figure 5) while the culture medium container is set in the support section, and the inside of the recess (e.g., recess 41a in Figure 5) can be kept constantly filled with disinfectant. Therefore, the connection section (e.g., the cap 23 of the connection section 40 in Figure 3) can be kept in a sterilized / disinfected state at all times.

[0090] (7) The culture medium container (for example, culture medium container 21 in Figures 3 and 4) A needle (for example, needle part 44 in Figure 9, etc.) A needle container for housing the needle (for example, the cup portion 45 in Figure 9), It has, The needle container (for example, the cup portion 45 in Figure 9) contains a sterilizing solution into which the needle (for example, the needle portion 44 in Figure 9) is immersed. As a result, when the tip of the needle (e.g., the needle portion 44 in Figure 9) is inserted into the cap, at least the needle (e.g., the needle portion 44 in Figure 9) is connected in a sterile state, immersed in the disinfectant 48 within the needle container (e.g., the cup portion 45 in Figure 9), thus preventing the entry of bacteria from the connection part, including the needle. In other words, bacterial contamination during container replacement can be prevented. [Explanation of symbols]

[0091] A...On-site culture system, D...Enclosure, S...Consumables sales business, E...End user, 1...Main unit, 2...Replacement equipment, 10...Control unit, 11...Blower, 12a, 12b...Pressure regulator, 13a, 13b...Flow meter, 14...Solenoid valve, 15a to 15i...Tube connector, 16...Membrane filter for air sterilization, 17, 18...Tubing pump, 19...Compressor, 21...Culture medium container, 23...Cap, 31...Culture vessel, 33...Silicone tubing, 35...Overflow tubing, 36...City water supply Tube for use, 37...Tube for culture medium outlet, 38...Tube for aeration air supply, 39...Capsule filter, 40...Connection part, 41...Container receiving part, 42...Needle connector, 43...Base part, 44...Needle part, 45...Cup part, 46...Cap, 47...Hook for silicone tube connection, 71...Hanging part, 72...Culture medium storage part, 73...Hanging hook hole, 74...Sterile water culture medium inlet, 75...Culture medium inlet, 76...Overflow outlet, 77...Aeration port, 78...Culture medium outlet, 111...Top lid seal, 112...Wooden cotton

Claims

1. In a microbial culture exchange device that functions as an exchange component that is detachably installed on the main body of a device for culturing target microorganisms, A culture vessel for culturing the aforementioned target microorganism, A culture medium container for storing consumables to be replenished to the culture vessel in liquid form, A liquid delivery unit that delivers the consumables from the culture medium container to the culture vessel, It is provided in a state that has been sterilized by gamma rays in advance. A replacement device for microbial culture.

2. The culture vessel has a bag-like shape. The exchange device for microbial culture according to claim 1.

3. The culture vessel has a tube connector for connecting other tubes to the infusion pack. The exchange device for microbial culture according to claim 2.

4. The culture medium container is made of a container that is expandable. The exchange device for microbial culture according to claim 1.

5. The culture medium container is The container body and A liquid outlet for dispensing the culture medium contained within the container body, A cap having a threaded portion that is screwed into the liquid outlet and a recess formed on the back side of the threaded portion, The recess in the cap contains absorbent cotton soaked in disinfectant solution, Equipped with, The exchange device for microbial culture according to claim 1.

6. The culture medium container is provided with a support part, The aforementioned support portion is A recess is filled with a disinfectant solution and houses the culture medium extraction section of the culture medium container so as to be immersed in the disinfectant solution, The disinfectant liquid supply section is provided, which protrudes radially from the edge surrounding the recess, Equipped with, The exchange device for microbial culture according to claim 1.

7. The culture medium container is Needle and, A needle container for housing the needle, It has, The needle container contains a disinfectant solution in which the needle is immersed. The exchange device for microbial culture according to claim 1.

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