JP2008161132A - Microorganism-culturing apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a compact microorganism-culturing apparatus of simple construction, capable of degerming, heating/making a stirring culture of a culture liquid, and lower in contamination risk than ever. <P>SOLUTION: The microorganism-culturing apparatus comprises an openable pressure-resistant closed vessel composed of a culture tank and a lid closing the culture tank, and a spawn injection part for sterilely injecting the culture tank with spawns. A culture liquid charged into the above closed vessel prior to making a culture is heated to its sterilizing temperature, and during making the culture, the temperature of the culture liquid is measured and the culture liquid is put to feedback heating control to a predetermined temperature. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a microorganism culture apparatus that is compact, can be heated and stirred with a household power source, and can grow only the target bacteria without contamination of the target microorganisms even outdoors.

  Conventional culturing apparatuses that require heating are used for industrial and research purposes, are large-sized, and sterilization is performed by a high-pressure steam sterilization method (120 ° C., about 2 atm, 15 to 20 minutes). This requires a large boiler or a commercially available high-pressure steam sterilizer. Assembling is difficult and expensive.

  Furthermore, in culture, the preparation of the medium and the introduction into the culture tank, the culture of the inoculum and the introduction into the culture tank must be performed aseptically. Furthermore, it is necessary to ensure aseptic conditions during culture. These operations require specialized knowledge and aseptic environmental equipment, and are difficult for farmers and ordinary households.

  Microbial culture apparatuses that can be cultivated outdoors are mostly sterilized by ultraviolet irradiation and administration of a bactericide. In such a sterilization method, germs (unintended microorganisms) cannot be sterilized in gaps where ultraviolet rays cannot be irradiated and gaps where the bactericide solution does not contact, and contamination has frequently occurred.

  In addition, in conventional culture tanks intended for outdoor operation, sterilization after adding the medium was not possible. This is because the effect of ultraviolet rays is not sufficiently exhibited due to the turbidity of the medium, and the target microorganism is sterilized, so that a bactericidal agent cannot be introduced.

  For this reason, the culture medium was prepared using a bypass tube in a culture tank in which a concentrated solution was prepared, sterilized, and sterilized water was collected. Accordingly, there are many cases in which only a target microorganism cannot be cultured when a person who has a high risk of contamination and is a general-purpose machine but does not actually have specialized knowledge works.

  As a compact microorganism culture apparatus capable of sterilization and culture, Patent Document 1 discloses a culture tank for holding a culture solution for culturing cells, a gas supply means for supplying gas into the culture solution, and stirring for stirring the culture solution. Means, a culture medium supply means for supplying a new culture medium to the culture tank, a culture medium discharge means for discharging the waste culture liquid from the culture tank, and an electromagnetic induction heating means, wherein the culture tank is provided with the electromagnetic induction A temperature measuring means for measuring the temperature of the culture solution in the culture tank; and the electromagnetic induction heating means based on the temperature measured by the temperature measuring means. A culture apparatus characterized by comprising control means for controlling and adjusting the amount of heating has been disclosed.

Further, as a sterilization method, Patent Literature 1 discloses a sterilization method for the culture apparatus, in which a culture solution is introduced into the culture tank, and the gas supply unit, the stirring unit, the culture solution supply unit, and the culture solution discharge unit. In the state where the culture device including the above is assembled, the control means is operated based on the sterilization conditions verified in advance, and the induction heating unit is heated by controlling the electromagnetic induction heating means. A method for sterilizing a culture apparatus characterized by sterilizing a culture solution and a culture instrument assembled in the culture tank has been disclosed.
JP 2006-129765 A

  However, the culture apparatus described in Patent Document 1 has a gas supply means for supplying gas into the culture solution and an agitation means for stirring the culture solution, that is, a stirring blade 17b, which is complicated and has a risk of contamination. It is.

  Further, although a specific method for inoculating the inoculum is not disclosed, from the description in Paragraph 0024 that the culture is introduced into the culture solution 11 and cultured, The lid is opened and the inoculum is introduced. Therefore, despite the sterilization of the inside of the culture tank, the culture tank is eventually opened when the bacteria are added, and the risk of contamination cannot be avoided.

  Accordingly, an object of the present invention is to provide a microorganism culturing apparatus that can sterilize, heat, and stir culture medium with a compact and simple configuration and has a lower risk of contamination.

  In order to solve the above problems, the present invention is connected to the open / close pressure-resistant airtight container 2 composed of a culture tank 2a and a lid 2b for sealing the culture tank 2a, and the open / close pressure-resistant airtight container 2 to aseptically inoculate the inoculum. The inoculum is injected into the inside 2f of the culture tank 2a, and the culture solution 8 introduced into the open / close pressure-resistant airtight container 2 is heated to the sterilization temperature before the culture, and the temperature of the culture solution 8 is maintained during the culture. , And the culture medium 8 is feedback-heat controlled to a predetermined temperature, the structure of the microorganism culture apparatus, the nut body 10a, the filler 10b filled in the inner space of the nut body 10a, and the filler After filling the nut body 10a, the nuts made of the first film 10c and the second film 10d, which are attached to the top and bottom of the nut body 10a and seal the filler 10b, are sterilized, and aseptically packaged 1 The inoculum 11 is filled in the cylinder of a sterilized syringe consisting of a cylinder 9a, a piston 9b, and a socket 9c fitted to the tip of the cylinder 9a. Thereafter, the outside of the socket 9c is covered with a film 9d and then aseptically sealed in a packaging bag 9e, and the inoculum injection part is provided with an open / close pressure-resistant sealed container 2 A connection pipe 4g connected to the connection pipe 4g, a valve 4d connected to the connection pipe 4g, an introduction pipe 4a connected to the valve 4d, and a sterilization nut 10 connected to the introduction pipe 4a. After being introduced into 2a, the open / close pressure-resistant airtight container 2 is sealed, the valve 4d is opened, and when the inoculum 11 is injected, the valve 4d is opened. A structure of the microorganism culture apparatus characterized in that the inoculum 9 containing the inoculum 11 is inserted into the upper part of the sterilizing nut 10, the inoculum 11 is injected into the open / close pressure-resistant sealed container 2, and then the valve 4d is closed to culture the microorganism. And the stirrer 7 is placed in the culture tank 2a, and the culture solution 8 is stirred by the stirrer 6 arranged below the bottom of the culture tank 2a.

  Since this invention is the above structure, the following effects are acquired. First, because it is a compact configuration, it can be easily transported, and can be used as a household 100V DC charger, so it can be used for farming, gardening at home, and outdoors. Use is also possible.

  Secondly, the inside of the open / close pressure-resistant sealed container is heated with an electric heating machine, and the diluted medium can be sterilized at a high temperature and can be sterilized under conditions close to the high-pressure steam sterilization method. There is no worry about nations. In addition, inoculation of the inoculum is performed by injecting the inoculum prepared by aseptically adjusting only the target inoculum from the inoculum injection section with a syringe, setting a disposable aeration filter during culture, and letting air naturally in the interior. Therefore, the risk of contamination is low even without specialized knowledge and aseptic environmental equipment. As a general-purpose machine, it is sufficient for farmers and general households to grow and propagate the target microorganisms without contamination.

  Thirdly, since the medium can be sterilized after preparation of the medium, the medium can be used in the form of powder instead of the concentrated liquid, so that problems such as quality deterioration after the concentrated liquid preparation does not occur.

  For the purpose of providing a versatile microbial culture apparatus that can be used in farmhouses and general households to prevent contamination, a culture tank 2a in which a stirring bar 7 is placed, and a pressure regulating apparatus on the 2f side inside the culture tank 2a Opening and closing pressure resistance comprising a lid 2b for sealing the culture tank 2a provided with the filters 17 and 18 for covering the ventilation holes of the float safety device 2q of 2c and before sterilization and the inoculum injection part 4 for aseptically adding the inoculum 11 A sealed container 2, a temperature sensor 5a for measuring the temperature of the culture solution 8 in the culture tank 2a, and a temperature control unit 5 for controlling the sterilization temperature and the culture temperature to a set temperature based on temperature information from the temperature sensor 5a; The stirrer 7 is rotated to stir the culture solution 8 and the stirrer 6 and the timer 3j for setting the sterilization time. A heating wire heater 3c for heating the container 2 and an electric heating type heater 3 on which the culture tank 2a is placed. The inoculum injection part 4 is connected to a switching tube 4g connected to the open / close pressure-resistant sealed container 2; A valve 4d connected to the connecting pipe 4g, an introduction pipe 4a connected to the valve 4d, a nut body 10a connected to the introduction pipe 4a, a filler 10b filled in the internal space of the nut body 10a, and the filling After filling the material 10b, the first film 10c is attached to the top and bottom of the nut body 10a to seal the filling material 10b, and the sterilized nut 10 is formed by sterilizing the nut made of the second film 10d. After charging, the open / close pressure-resistant airtight container 2 is sealed, the valve 4d is opened, and when the inoculum 11 is injected, the valve 4d is opened. Furthermore, the inoculum 11 is filled into the inside of the cylinder 9a of a sterilized syringe consisting of the cylinder 9a, the piston 9b, and the socket 9c fitted to the tip of the cylinder 9a, and then the outside of the socket 9c is covered with a film 9d. Realized by the configuration of the microorganism culture apparatus 1 characterized by stabing the syringe 9 containing the inoculum 11 and injecting the inoculum 11 into the open / close pressure-resistant airtight container 2 and then closing the valve 4d to culture the microorganism. did.

  Hereinafter, a microorganism culture apparatus according to the present invention will be described in detail with reference to the accompanying drawings. FIG. 1 is a perspective view of a microorganism culture apparatus according to the present invention. A microorganism culture apparatus 1 according to the present invention includes an open / close pressure-resistant sealed container 2 for culturing a target microorganism, a temperature control unit 5 for controlling the temperature in the open / close pressure-resistant sealed container 2, and the culture tank 2a. It comprises an electric heating heater 3 for heating the pressure-resistant sealed container 2.

  The open / close pressure-resistant sealed container 2 includes a culture tank 2a and a lid 2b that seals the culture tank 2a, and is connected to the open / close pressure-resistant sealed container 2 to inoculate inoculum into the culture tank 2a aseptically. Part 4 is provided.

  The open / close pressure-resistant airtight container 2 is provided with a hole in a lid of a pressure cooker equipped with a pressure regulating device 2c for regulating the pressure in a generally circulating pan, and a temperature sensor 5a is inserted and fixed, and an inoculum injection portion 4 may be connected. As the pressure cooker, a stainless steel pressure cooker, for example, a two-handed pressure cooker 8L (product number 601162), 10L (product number 601179) manufactured by Ito Aluminum Industry Co., Ltd. is suitable. However, the pan volume can be appropriately selected depending on the culture amount of the microorganism.

  The temperature control unit 5 controls the sterilization temperature and the culture temperature to the set temperature based on the temperature information from the temperature sensor 5a that measures the temperature of the culture solution in the culture tank 2a.

  The electric heating machine 3 mounts the culture tank 2a, heats the culture solution put in the open / close pressure-resistant airtight container before culture to the sterilization temperature, and measures the temperature of the culture solution with the temperature sensor 5a during the culture. Then, feedback heating control of the culture solution to a predetermined culture temperature is performed.

  FIG. 2 is a partially broken side view of the microorganism culture apparatus according to the present invention. The open / close pressure-resistant airtight container 2 is fixed to the installation part 3b on the electric heating type heater 3 by an edge 3a and placed in close contact therewith. Therefore, it is desirable that the upper surface of the installation portion 3b of the electric heating heater 3 has a shape into which the shape of the bottom of the open / close pressure-resistant sealed container 2 is fitted.

  The temperature sensor 5a is inserted so that the temperature of the culture solution 8 can be measured. Of course, the insertion hole of the temperature sensor 5a is sealed so that the internal pressure does not leak. The stirrer 7 may be put in the culture solution 8 inside the culture tank 2a before the culture solution 8 is sterilized. The stirrer 7 is an instrument in which a magnet is embedded and rotated by a stirrer 6 to stir the liquid, and has heat resistance at a sterilization temperature of 120 ° C.

  The electric heating machine 3 is partially shown as a perspective view. Inside the electrothermal heater 3, a so-called stirrer 6 using a magnet is incorporated, and the stirrer 7 is rotated so that the culture solution 8 can be stirred as needed during the culture. For example, Oriental Motor Co., Model No. IK6A-AWJ650 can be used as the stirrer 6. Further, a ventilation hole 3d is provided for exhausting heat generated by the rotation of the stirrer 6 with a fan.

  The temperature (sterilization temperature) in the culture tank 2a when there is no culture solution 8 and the temperature (culture temperature) of the culture solution 8 are measured by the temperature sensor 5a, and the inner surface of the installation part 3b is controlled by the control system in the electric heating machine 3 Heating is performed by a heating wire heater 3c installed in the apparatus, and the temperature in the culture tank 2a is feedback-heat controlled to a predetermined temperature.

  FIG. 3 is a plan view of the microorganism culture apparatus according to the present invention. The culture tank 2a and lid 2b of the open / close pressure-resistant airtight container 2 used here are a two-handed pressure cooker 10L (part number 601179) manufactured by Ito Aluminum Industry Co., Ltd. Details of the adjustment of the internal pressure of the culture tank 2a are in the instruction manual. The pressure adjusting device 2c is composed of a weight, a nozzle, and a nozzle filter. Excess pressure is discharged from the nozzle, and the pressure during boiling is maintained at 98 kilopascals by the action of the weight and the nozzle.

  The safety device 2p at the base of the left handle 2d operates when the pressure adjusting device 2c and the float safety device 2q do not operate, and when the pressure in the culture tank 2a reaches 200 kilopascals, and the steam in the culture tank 2a. It is a safety device that discharges water.

  The float type safety device 2q at the base of the right handle 2e floats when there is pressure in the culture tank 2a, sinks when the pressure approaches 0, and the nozzle of the pressure adjusting device 2c is clogged with the contents This is a safety device that discharges steam when the internal pressure reaches 150 kilopascals, and keeps the pressure in the culture tank 2a below 150 kilopascals. In the present invention, as long as the temperature of the internal culture solution 8 can reach 120 ° C. and a valve is attached, the present invention is not limited to these safety devices.

  The inoculum injection part 4 is connected to the lid 2b here, but may be attached to the side surface of the culture tank 2a. Similarly, the temperature control unit 5 may be inserted through the temperature sensor 5a from the side surface of the culture tank 2a. The sterilization nut 10 connected to the upper end of the valve 4d is covered with a second film 10d whose upper surface seals the inside of the culture tank 2a. That is, a state before inoculating the culture tank 2a is shown.

  FIG. 4 is a plan view of the microorganism culturing apparatus according to the present invention with the lid removed. FIG. 5 is a bottom view of the lid of the microorganism culture apparatus according to the present invention. The culture solution 8 is put into the inside 2f of the culture tank 2a, sealed, and the target microorganism is heated and cultured.

  In order to seal the culture tank 2a with the lid 2b, the lid 2b is pressed against the upper edge of the culture tank 2a, and the left and right fixing grooves 2i and 2j provided in the left and right fixing portions 2g and 2h of the culture tank 2a are inserted into the left and right sides of the lid 2b. The left and right rods 2n and 2o provided on the handles 2d and 2e are rotated and fitted and fixed. At this time, the outer protrusion 2r of the culture tank 2a and the inner protrusion 2m of the lid 2b are engaged and fixed, and the open / close pressure-resistant sealed container 2 is sealed by the packing 2s installed on the inner protrusion 2m of the lid 2b. The sealing means is not limited to this as long as it is a pressure resistant container whose lid can be opened and closed. It is only necessary to maintain hermeticity so that the internal 2f temperature can withstand 120 ° C.

  An inoculum injection part 4 is connected to the lid 2b, and an introduction path 2l penetrating from the valve 4d passes through the inside 2f of the culture tank 2a. Further, a temperature sensor insertion hole 2k through which the temperature sensor 5a is inserted is formed, and the temperature sensor 5a is inserted into the culture tank 2a.

  Here, the microorganism culturing apparatus 1 of the present invention prevents contamination during culture by not forcing a gas such as outside air into the culture tank 2a. Further, all the filters are attached to the holes communicating with the outside air in the inside 2f of the culture tank 2a.

  Since the lid 2b used in the present invention is a two-handed pressure cooker 10L manufactured by Ito Aluminum Industry Co., Ltd., a pressure adjusting device 2c (dotted line in FIG. 4) with outside and in / out of air, and a culture tank for a float type safety device 2q 2a The inside 2f side part is covered with filters 17 and 18.

  The filters 17 and 18 are attached to all the holes communicating with the outside air before the sterilization process of the culture solution 8. The material is not particularly limited as long as it has excellent air permeability with a pore diameter of about 0.1 to 0.7 μm, has heat resistance even at a sterilization temperature (120 ° C.), and is waterproof. For example, Milli-Seal manufactured by Millipore (having a pore diameter of 0.45 μm) can be used, and an adhesive surface formed on one surface can be easily attached to a hole that communicates with the outside air. .

  FIG. 6 is a plan view of an electrothermal heater constituting the present invention. The electric heater 3 incorporates a heating wire heater 3c in the lower part of the installation part 3b where the open / close pressure-resistant sealed container 2 is installed. The heating wire heater 3c is disposed so as to surround the stirrer 6 inside the edge 3a to which the open / close pressure-resistant airtight container 2 is fitted and fixed. In addition, as long as the heating wire heater 3c is in contact with or close to the inner surface of the installation portion 3b, the wiring is not particularly limited.

  FIG. 7 is an enlarged perspective view of an inoculum injection portion of the microorganism culture device according to the present invention. The inoculum injection unit 4 includes a connecting pipe 4g connected to the lid 2b of the open / close pressure-resistant airtight container 2, a valve 4d connected to the connecting pipe 4g, an introduction pipe 4a connected to the valve 4d, and an introduction pipe 4a. It consists of a connected sterilization nut 10.

  The connecting pipe 4g is inserted through the introduction path 2l, which is a path through which the inoculum is injected, and is connected to the culture tank 2a. The connecting pipe 4g is connected to the lid 2b by welding or the like, but when the strength is weak, a reinforcing member 4h, which is a triangular plate, is welded to the side surface of the connecting pipe 4g and the surface of the lid 2b. It is good to reinforce 4g so that it may resist resistance from the side.

  The valve 4d is a handle type screw-in stop valve here, and is screwed onto the connecting pipe 4g and the introduction pipe 4a with the nut portions 4c and 4e of the valve 4d. However, the valve 4d is a path through which the inoculum 111 is injected into the culture tank 2a. The connection method and type are not particularly limited as long as a certain introduction path 21 can be opened and closed and can withstand the heating temperature and the heating pressure.

  For example, it may be connected to the culture tank 2a and the lid 2b by a flange shape or welding, and gate valves, globe valves, ball valves, butterfly valves, needle valves, stop valves, check valves, etc. can be used.

  The introduction tube 4a is connected to the valve 4d, penetrates the sterilization nut 10 through the first film 10c on the lower surface, and the distal end portion 9f of the syringe 9 is inserted. The distal end portion 9f of the syringe 9 may be a needle or a socket. The socket is a tip attached to the tip of a micropipette or the like.

  The syringe 9 includes a cylinder 9a in which an inoculum 11 is placed, a piston 9b that pushes out the inoculum 11, and a tip 9f that is fitted to the tip of the cylinder 9a. The inoculum 11 is aseptically filled into the cylinder 9a so as not to contain bacteria other than the cultured bacteria.

  In order to aseptically put the inoculum 11 into the open / close pressure-resistant sealed container 2, for example, in the case of the configuration of FIG. 7, after the culture solution 8 is introduced into the culture tank 2a, the open / close pressure-resistant sealed container 2 is sealed. Then, the valve 4d is opened, the valve 4d is opened when the inoculum 11 is injected, the syringe 9 containing the inoculum 11 is inserted into the upper part of the sterilizing nut 10, and inserted into the introduction tube 4a. 2l is injected into the open / close pressure-resistant airtight container 2, and then the valve 4d is closed to culture the microorganism.

  FIG. 8 is a cross-sectional view showing a state before the inoculum injection part is mounted. FIG. 8A shows an inoculum set 19 prepared by aseptically adjusting the inoculum 11 to be introduced into the microorganism culture apparatus 1 according to the present invention. FIG. 8B shows a disposable sterilization nut 20 for aseptically providing the sterilization nut 10 that constitutes the inoculum injection part 4 of the microorganism culture apparatus 1 according to the present invention.

  The inoculum set 19 includes a syringe 9, an inoculum 11 filled in the cylinder 9 a of the injector 9, and a packaging bag 9 e for aseptically packaging the syringe 9 filled with the inoculum. The syringe 9 includes a cylinder 9a for placing the inoculum 11 therein, a piston 9b for extruding the inoculum, and a tip 9f fitted to the tip of the cylinder 9a. The tip portion 9f is composed of a socket 9c fitted to the tip of the cylinder 9a and a film 9d that covers the outer periphery of the socket 9c so that inoculum does not leak.

  The socket 9c is a portion through which the sterilizing nut 10 is penetrated, and a tube having a thinner heat-resistant tip that can withstand sterilization such as an autoclave, for example, a needle, or a micropipette usually used in chemical or biological experiments. There are chips (yellow chips, blue chips, etc.) that are mounted on.

  The film 9d is a thin film that covers the outer surface of the socket 9c and is wound so that the inoculum 11 inside the syringe 9 does not leak. If it is excellent in adhesiveness and waterproofness and can seal the syringe 9, it will not specifically limit. For example, there is Parafilm (registered trademark).

  The procedure for creating the inoculum set 19 is as follows. In a sterile area such as a clean bench, the sterilized socket 9c is fitted to the tip of a sterilized plastic syringe, and a pre-cultured inoculum 11 is sucked from the tip of the socket 9c by a predetermined amount. The entire outer surface of the socket 9c is covered with a film 9d so that the inoculum 11 inside the cylinder 9a does not leak, and finally, the syringe 9 containing the inoculum 11 is put in a plastic packaging bag 9e through which microorganisms, molds, etc. do not permeate, sealers, etc. Heat seal and seal.

  Thus, since the syringe 9 containing the inoculum 11 does not come into contact with other bacteria and the inoculum 11 can be stored without causing contamination, the inoculum can be stored in the microorganism culture apparatus 1 according to the present invention as necessary. This is convenient.

  The disposable sterilization nut 20 is formed by aseptically sealing and packaging the sterilization nut 10 with a packaging bag 10e or the like. The sterilizing nut 10 includes a nut body 10a, a filler 10b filled in the inner space of the nut body 10a, and a first filler that is applied to the top and bottom of the nut body 10a after being filled with the filler 10b to seal the filler 10b. The sterilized nut 10 sterilized from the nut made of the film 10c and the second film 10d is sterilized and then aseptically sealed in the packaging bag 10e.

  The filler 10b is a plastic material such as a polyethylene sheet, and is a sponge-like or foam-like material. The filler 10b pushes in a volume equal to or larger than the internal space of the nut body 10a, and the first and second films 10c, Sealed at 10d.

  The first film 10c and the second film 10d are metallic thin sheets coated with an adhesive on one side, and after the filler 10b is pushed in, the first film 10c and the second film 10d are attached to the edge of the nut body 10a. For example, an aluminum tape S (model number J3230) manufactured by Nitoms Corporation is suitable because it is inexpensive and easily available.

  9 and 10 are cross-sectional views for explaining a procedure for mounting the inoculum injection part. FIG. 12 is a cross-sectional view showing a state after the inoculum injection part is mounted.

  Hereinafter, a method for assembling the inoculum injection unit 4 will be described with reference to FIGS. 9 to 11. First, before sterilization and before sterilization, the sterilizing nut 10 shown in FIG. 9 (A) is screwed into the upper end of the introduction pipe 4a connected to the nut portion 4c of the valve 4d shown in FIG. 9 (B). Therefore, the 1st film 10c must be a raw material of the intensity | strength torn at this time.

  FIG. 10B shows a state after the sterilization nut 10 is screwed into the introduction pipe 4a. After the screwing, the filler 10b press-fitted into the nut body 10a is further compressed and exists between the second film 10d and the introduction pipe 4a.

  With the sterilization nut 10 screwed into the introduction pipe 4a, the valve 4d is opened, and the culture solution 8 and the inside of the open / close type pressure-resistant sealed container 2 are heated through the culture tank 2a, the introduction path 2l, inside the valve 4d, and the pipe line 4i. Sterilize. In the microorganism culture apparatus 1 according to the present invention, heating is performed by feedback control of the electric heating heater 3, the heating wire heater 3 c, and the temperature control unit 5. As a result, the introduction path 21 of the connection pipe 4g, the inside of the valve 4d, and the pipeline 4i of the introduction pipe 4a are also sterilized.

  After sterilization of the culture solution 8 and the open / close pressure-resistant sealed container 2, the inoculum 11 containing the syringe 9 prepared in advance shown in FIG. 10 (A) is applied to the second film 10d on the upper surface of the sterilization nut 10 in FIG. 10 (B). The tip 9f of the syringe 9 is pierced.

  FIG. 11 and FIG. 12 are cross-sectional views illustrating an aseptic method for inoculating inoculum. As shown in FIG. 11, the second film 10d is torn by piercing the second film 10d on the upper surface of the sterilizing nut 10 with the tip 9f of the syringe 9. Further, by inserting the distal end portion 9f into the pipe 4i, the second film 10d, the filler 10b, and the distal end portion 9f are rubbed, and the film 9d covering the outer surface of the socket 9c is torn and peeled off from the socket 9c. A hole 9g at the tip of the socket 9c is exposed inside the conduit 4i of the introduction tube 4a.

  As shown in FIG. 12, the tip of the tip 9f is inserted as far as possible into the pipe 4i, the piston 9b of the syringe 9 is pushed into the cylinder 9a, and the inoculum 11a is put into the pipe 4i, the valve 4d, and the introduction path. 2l and aseptically injected into the inside 2f of the culture tank 2a. FIG. 12 is a cross-sectional view of the state of FIG. After inoculating the inoculum 11 into the culture tank 2a, the valve 4d is closed. Then, if necessary, heating, the stirring bar 7 is rotated, the culture solution 8 is stirred, and cultured for a predetermined time. After the cultivation is completed, the lid 2b is opened, and the cultured microorganisms are sprayed on the fields, flower beds, vegetable gardens and the like.

  By aseptically adjusting the inoculum 11 to the configuration, assembly, and syringe 9 of the inoculum injection unit 4, the risk of contamination when the inoculum 11 is injected into the culture tank 2a is extremely reduced.

  FIG. 13 is a circuit diagram of a microorganism culture apparatus according to the present invention. FIG. 14 is a diagram showing an example of the operation panel of the present invention.

  The circuit 13 of the microorganism culturing apparatus is a circuit corresponding to the operation panel 3f of FIG. 14, and is roughly divided into a sterilization circuit 14 and a culture circuit 15. Here, the sterilization temperature was 120 ° C., and the culture temperature was 30 ° C. The plug 12 can be connected to a household 100V to supply electricity to the circuit. The power supply is turned on and off with a switch of the main power supply 3g. When the power is turned on, the power lamp 3h is lit.

  The sterilization temperature and the culture temperature are measured by the temperature sensor 5a of the temperature control unit 5, and the temperature of the culture solution 8 in the culture tank 2a is measured, and feedback control is performed by the 120 ° C temperature adjuster 5b and the 30 ° C temperature adjuster 5c, respectively. Is done.

  The sterilization temperature can be arbitrarily set on the sterilization temperature setting panel 3i. Generally, 120 degreeC and 20 minutes which kill the heat-resistant microbes which form spores, such as a Bacillus subtilis, are set. Further, the culture time can be arbitrarily set on the culture time setting panel 3o according to the characteristics of the microorganism to be cultured.

  The 120 ° C. and 30 ° C. temperature controllers 5b and 5c are temperature control devices using a bimetal thermostat, and feedback control the temperature in the culture tank 2a to the set temperature together with the temperature sensor 5a and the electric heater 3c.

  When the culture solution 8 or predetermined water and powder for culture solution are put into the culture tank 2a and the sterilization start switch 3k is pressed, the sterilization lamp 3l is turned on and the stirrer 6 rotates the stirring bar 7 While stirring the culture solution 8, the predetermined sterilization temperature and time are maintained, and the culture solution 8 is sterilized.

  Next, after the set time of the timer 3j elapses, the sterilization lamp 3l is turned off, and the sterilization end lamp 3n for displaying the end of sterilization is turned on. When trouble occurs, the sterilization stop switch 3m can be turned on to forcibly stop the sterilization process.

  Then, it is left for a whole day and night, and when the culture solution 8 is cooled to about room temperature, the inoculum 11 is injected into the open / close pressure-resistant airtight container 2 by the above-mentioned means. After the injection of the inoculum 11, the valve 4d is closed, the culture start switch 3p is turned on, and the culture is started.

  During the culture, the culture lamp 3q is turned on, the temperature of the culture solution 8 is measured in real time by the temperature sensor 5a, and feedback control is performed to a predetermined temperature (here, 30 ° C.) with a 30 ° C. temperature controller. The heating temperature is controlled by the heating wire heater 3c. At the same time, the fan 3c is rotated to cool the interior of the electric heating machine 3. Moreover, the stirrer 6 stirs and stirs the culture solution 8 during culture. Stirrer 6 can be a separate system, and stirring can be arbitrarily selected as necessary. When trouble occurs, the culture process can be forcibly terminated by the culture stop switch 3r.

It is a perspective view of the microorganism culture apparatus which is this invention. It is a partially broken side view of the microorganism culture apparatus which is this invention. It is a top view of the microorganism culture apparatus which is this invention. It is a top view except the lid | cover of the microorganism culture apparatus which is this invention. It is a cover bottom view of the microorganism culture apparatus which is this invention. It is a top view of the electric heating type heater which constitutes the present invention. It is an expansion perspective view of the inoculum injection part of the microorganism culture apparatus which is this invention. It is sectional drawing which shows the state before mounting | wearing of an inoculum injection part. It is sectional drawing explaining the mounting | wearing procedure of an inoculum injection part. It is sectional drawing explaining the mounting | wearing procedure of an inoculum injection part. It is sectional drawing explaining the aseptic injection method of an inoculum. It is sectional drawing explaining the aseptic injection method of an inoculum. It is a circuit diagram of the microorganism culture apparatus which is this invention. It is a figure which shows an example of the operation panel of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microbe culture apparatus 2 Opening-closing pressure-resistant airtight container 2a Culture tank 2b Lid 2c Pressure adjustment apparatus 2d Left handle 2e Right handle 2f Inside 2g Left fixed part 2h Right fixed part 2i Right fixed groove 2j Left fixed groove 2k Temperature sensor insertion hole 2l Introduction Road 2m Inner projection 2n Left side 2o Right side 2p Safety device 2q Float type safety device 2r Outer projection 2s Packing 3 Electric heating heater 3a Edge 3b Heating wire heater 3d Ventilation hole 3e Fan 3f Operation panel 3g Main power source 3h Power source Lamp 3i Sterilization temperature setting panel 3j Timer 3k Sterilization start switch 3l Sterilization lamp 3m Sterilization stop switch 3n Sterilization end lamp 3o Incubation time setting panel 3p Incubation start switch 3q Incubation lamp 3r Incubation stop switch 3s Heater energization control relay switch 4 Inoculum injection 4a Inlet pipe 4b Inlet 4c Nut Part 4d Valve 4e Nut part 4g Connecting pipe 4h Reinforcing material 4i Pipe line 4j Filter 5 Temperature control unit 5a Temperature sensor 5b 120 ° C temperature regulator 5c 30 ° C temperature regulator 6 Stirrer 7 Stirrer 8 Culture solution 9 Syringe 9a Cylinder 9b Piston 9c Socket 9d Film 9e Packaging bag 9f Tip 9g Hole 10 Sterilization nut 10a Nut body 10b Filler 10c First film 10d Second film 10e Packaging bag 11 Inoculum 11a Inoculum 12 Plug 13 Microbial culture apparatus circuit 13a Motor control relay 13b Sterilization Medium heater overheat relay 13c Sterilization time setting timer relay 13d Sterilization end relay 13e Culture heater overheat relay 13f Culture time setting timer relay 13g Empty prevention switch 14 Sterilization circuit 15 Culture circuit 16 Timer circuit 1 7 Filter 18 Filter 19 Inoculum Set 20 Disposable Sterilization Nut

Claims (5)

  A culture tank and an open / close pressure-resistant airtight container composed of a lid for sealing the culture tank, and an inoculum injection section that is connected to the open / close pressure-resistant airtight container and aseptically injects the inoculum into the culture tank. The microorganism culture apparatus is characterized in that the culture liquid put in the open / close pressure-resistant airtight container is heated to a sterilization temperature, the temperature of the culture liquid is measured during the culture, and the culture liquid is feedback-heat controlled to a predetermined temperature. .   After sterilizing the nut made of the nut body, the filler filled in the inner space of the nut body, the first film and the second film that are stuck on the top and bottom of the nut body and sealed with the filler after filling the filler, A disposable sterilized nut set, which is aseptically sealed in a packaging bag.   The inoculum is filled in the cylinder of a sterilized syringe consisting of a cylinder, a piston, and a socket fitted to the tip of the cylinder, and then the outside of the socket is covered with a film and then aseptically sealed in a packaging bag. Inoculum set characterized by that.   The sterilization nut according to claim 3, wherein the inoculum injection portion is connected to a switching pipe pressure-resistant airtight container, a valve connected to the connecting pipe, an introduction pipe connected to the valve, and a sterilization nut connected to the introduction pipe. After the culture solution is put into the culture tank, the open / close pressure-resistant airtight container is sealed, the valve is opened, the valve is opened at the time of inoculation of the inoculum, a syringe containing the inoculum is inserted on the sterilization nut, and the inoculum is 2. The microorganism culturing apparatus according to claim 1, wherein the microorganism is cultured by injecting into an open / close pressure-resistant sealed container, and then closing the valve.   The microorganism culture apparatus according to claim 1 or 4, wherein a stir bar is placed in the culture tank, and the culture solution is stirred with a stirrer disposed below the bottom of the culture tank.

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