JP2016086654A - Microorganism culture apparatus, microorganism culture system, and microorganism culture method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a microorganism culture apparatus, a microorganism culture system, and a microorganism culture method in which adjustment or control of a culture condition can be performed simply and easily and which can be used generally for isolation culture of microorganism in an environment.SOLUTION: A microorganism culture apparatus 4 can supply culture solution continuously to a porous body 1 in the state where a liquid level of the culture solution is formed in the vicinity of the surface of the porous body and slightly below the surface of the porous body. Therefore, a supply amount and a composition of the culture solution can be arbitrary changed on the surface of the porous body 1 while a microorganism is cultured. Therefore, types of microorganisms which can be cultured can be increased. Moreover, because continuous culture can be performed in the state where individual colonies are isolated, a useful microorganism can be individually isolated.SELECTED DRAWING: Figure 4

Description

  The present invention relates to an apparatus, system, and method for culturing microorganisms, and more particularly, to a microorganism culture apparatus, a microorganism culture system, and a microorganism culture method that can be generally used for separation and culture of microorganisms in the environment.

  In recent years, with the development of microbial engineering, substance production using useful microorganisms and environmental purification have been performed. On the other hand, when molecular phylogenetic analysis is performed on the basis of information on rRNA genes obtained from the environment, it is clear that there are enormous species of microorganisms on the earth. For example, it is estimated that more than 10,000 to 1,000,000 kinds of microorganisms exist in a sample of about 1 to 10 g of soil.

  In other words, it produces compounds that can be used in pharmaceuticals, foods, cosmetics, etc. in natural ecosystems such as rivers, lakes, seawater, soil and hot springs, and artificial ecosystems such as wastewater treatment, waste treatment, and composting. It is estimated that there are many unknown useful microorganisms. In other words, the microorganisms that are used are only a small part of the existing species, and therefore, there is an interest in obtaining useful microorganisms.

  However, since many kinds of microorganisms live together in such an ecosystem, it is necessary to collect (separate) target useful microorganisms from the complex microorganism group. In the case of industrial use, if the process is built using the microbial community in the natural environment as it is, it is difficult to control the stable production of useful compounds that can be used in pharmaceuticals, foods, cosmetics, etc. This is because it is desirable to use a specific microorganism that has been confirmed.

  Conventionally, target microorganisms are collected by pure culture of microorganisms. For this pure culture, a method for separating and culturing microorganisms since the Koch era is generally used. Specifically, it is a method of culturing microorganisms using a solid medium. First, an appropriate concentration of agarose is dissolved in a medium containing an organic or inorganic substrate that can be used by microorganisms, and poured into a petri dish to prepare a solid medium. Next, the microorganism group collected from the environment is appropriately diluted on the surface of the solid medium, smeared and cultured. If the dilution stage of the seeding source is adjusted, about 10 to 100 microbial colonies can be formed on the solid medium (agar plate surface smearing method). In place of agarose, gellan gum, silica gel or the like is also used as appropriate.

  These colonies are considered to be formed by growing one microbial cell on a solid medium. The purity of microorganisms can be improved by repeating a procedure in which one colony is dispersed with sterilized water and then smeared on a solid medium to form a colony again.

  However, there are many things that do not grow in the conventional pure culture using a solid medium, and it is known that about 1% of microorganisms that can be cultured currently exist in the environment. The reason is that the culture environment is a closed environment, so that excess microorganism-generated substances cannot be discharged out of the system. This is presumed to be because metabolites and signal factors produced by microorganisms accumulate, and microorganisms that are inhibited by them cannot grow. Another factor is that it is difficult to maintain the substrate concentration required for the growth of the target microorganism. For this reason, a new isolation culture technique different from the pure culture using the conventional agar plate is required.

  For example, Patent Document 1 discloses that after a solid medium mixed with an inoculum is sealed in a container in which a film having pores that permeate a liquid phase containing a nutrient source and the like and do not allow microorganisms to permeate is sealed, the container A technique for cultivating microorganisms by exposing the solid medium to the original growth environment of the microorganisms by installing in the environment is disclosed. As a result, attempts have been made to culture microorganisms that can only live in natural environments.

  Furthermore, in Patent Document 2, a large number of gel particles in which one microbial cell is embedded are generated, and the gel particles together with another gelling agent serving as a matrix pass through various chemical substances without passing through the microorganisms. A technique is disclosed in which microorganisms in gel particles are cultured by injecting into a tube formed of a film body that accepts water and allowing it to gel, and placing the tube in the environment.

  Further, in Patent Document 3, in order to separate and culture difficult-to-cultivate microorganisms that can only live in the presence of symbiotic microorganisms, a solid medium for culturing symbiotic microorganisms is provided, and a filter is provided on the solid medium via a filter. A culture system in which a solid medium for culturing difficult-to-cultivate microorganisms is stacked is disclosed. The filter between the solid media prohibits the movement of the microorganisms, but allows the products of the microorganisms to pass therethrough, so that the difficult-to-culture microorganisms can be separated and cultured in independent compartments, that is, without being mixed with the symbiotic microorganisms.

JP-T-2005-503171 JP 2010-41967 A JP 2012-175773 A

  However, the technique described in Patent Literature 1 has a problem that colonies of microorganisms formed by liquid phase flow generated in the solid medium may disappear, and colony formation tends to be unstable. In addition, colonies in which a plurality of microorganisms are mixed may be formed along with the liquid phase flow. That is, the probability that the formed colony is a single colony that is an aggregate of one microorganism is reduced, and it is difficult to culture (separate) a specific microorganism alone as in the above pure culture. Moreover, about the colony inside a solid medium, there existed a problem that it was difficult to isolate | separate and acquire only a single colony so that other microbial cells might not be mixed.

  Furthermore, the technique described in Patent Document 2 requires highly skilled operations for producing gel particles, and further, collecting them while confirming whether or not colonies are generated in minute gel particles. Exquisite and complicated work is required. For this reason, there has been a problem that the workload of the worker is high.

  Moreover, in the technique described in the said patent document 3, nutrient supply is performed to the agar culture medium by which microorganisms are culture | cultivated by the material exchange from a solid medium which stocks a nutrient source, or a concentration gradient. For this reason, there is a problem that the moving speed of the nutrient source or the like tends to be slow, and the nutrient source supply becomes insufficient, which may cause a culture failure of the microorganism. Furthermore, in the present technology, when the product of the microorganism is not sufficiently consumed by the symbiotic microorganism, the product is stored in the solid medium. In such a case, the problem that the growth of microorganisms may be inhibited by the product cannot be solved. Furthermore, since the culture is performed in a state where a plurality of layers of solid culture media are stacked, the process is complicated and complicated. In addition, in order to separate and culture a wide variety of microorganisms, it is necessary to set individual culture conditions suitable for each microorganism. However, in this technology in which culture is performed with a predetermined medium composition, the culture conditions can be freely adjusted. There is a problem that it is difficult to apply widely to culture of various microorganisms.

  The present invention has been made in order to solve the above-described problems, and can be easily and easily adjusted and controlled for culture conditions, and can be used for general-purpose separation and culture of microorganisms in the environment. An object is to provide a culture apparatus, a microorganism culture system, and a microorganism culture method.

  In order to achieve this object, the microorganism culturing apparatus according to claim 1 is an apparatus for cultivating microorganisms, has a pore configured to be permeable to liquid and communicated with the surface, and has pores on the upper surface portion. A culture chamber in which a porous body formed with a pore size smaller than that of a microorganism is disposed, a flow path for supplying and discharging liquid to the culture chamber, a holding unit for holding the porous body in the culture chamber, The liquid level formed by supplying the liquid from the flow path to the culture chamber is the same height as the upper surface of the porous body held by the holding unit or near the upper surface of the porous body and below the upper surface of the porous body. And adjusting means for adjusting the positional relationship between the porous body and the liquid surface, and the same height as the upper surface of the porous body or slightly lower while supplying liquid continuously or intermittently to the culture chamber In the state where the liquid level is formed at the position, The liquid is supplied to the porous body top surface through the pores of the porous body, in which culturing a microorganism on multi hole body top surface.

  Here, the adjusting means is for the relative positional relationship between the liquid surface and the upper surface of the porous body to be the above-described relationship, and by adjusting the height of the porous body, The positional relationship may be adjusted, or the positional relationship with the upper surface of the porous body may be adjusted by adjusting the height of the liquid surface. Moreover, you may adjust both positions, respectively.

  The microorganism culturing apparatus according to claim 2 is the microorganism culturing apparatus according to claim 1, wherein the holding portion includes a pedestal on which the porous body is placed, and a detachable engagement with the porous body on the pedestal. The adjusting means includes a column provided on the holding unit, to which the pedestal is detachably attached, and a fixture for fixing the column to the pedestal. The positional relationship between the upper surface of the porous body and the liquid surface is adjusted by changing the height of the pedestal using a support post.

  The microorganism culture apparatus according to claim 3 is the microorganism culture apparatus according to claim 1, wherein the holding portion is a base on which the porous body is placed, and a detachable engagement with the porous body on the base. The adjusting means includes a support provided on the holding portion, to which the pedestal is attached, and a fixture for fixing the pedestal so that the pedestal can be moved in the vertical direction with respect to the support. The positional relationship between the upper surface of the porous body and the liquid surface is adjusted by moving the pedestal in the vertical direction.

  The microorganism culture apparatus according to claim 4 is the microorganism culture apparatus according to claim 2 or 3, wherein the pedestal includes a through-hole through which the liquid supplied to the culture chamber passes in the thickness direction of the pedestal. Yes.

  The microorganism culture apparatus according to claim 5 is the microorganism culture apparatus according to any one of claims 1 to 4, wherein the porous body is a container in which a through-hole communicating between the inside and the outside of the container is formed. The container has a container, and the holding unit holds the container in the culture chamber.

  The microorganism culture apparatus according to claim 6 is the microorganism culture apparatus according to any one of claims 1 to 5, wherein the porous body has an upper surface formed of a gel.

  The microorganism culture apparatus according to claim 7 is the microorganism culture apparatus according to claim 6, wherein the entire porous body is formed of a gel.

  The microorganism culturing apparatus according to claim 8 is the microorganism culturing apparatus according to any one of claims 1 to 7, comprising a bottomed casing that opens upward, and a lid that closes the opening. An outer tank containing the culture chamber is provided with a gap between the inner wall of the housing and the flow path passes through the wall of the outer tank and is directly connected to the culture chamber.

  The microorganism culturing apparatus according to claim 9 is the microorganism culturing apparatus according to claim 8, wherein the outer tub is provided in a jig for fixing the lid body in a closed state and an opening portion of the housing. And a sealing material that prevents entry of outside air when the lid is fixed by the tool, and is configured to be hermetically sealed.

  The microorganism culturing apparatus according to claim 10 is the microorganism culturing apparatus according to claim 9, wherein an oxygen scavenger is provided in the internal space of the outer tank, and the inside of the outer tank is made an anaerobic atmosphere by the oxygen scavenger. .

  A microorganism culturing system according to an eleventh aspect includes a plurality of the microorganism culturing apparatuses according to any one of claims 1 to 10, a storage tank in which liquids supplied to the plurality of microorganism culturing apparatuses are stored, and storage thereof A supply pump for supplying a liquid stored in a tank to a flow path connected to each of the plurality of microorganism culture apparatuses; and a discharge pump for discharging the liquid from each of the microorganism culture apparatuses. Culture is performed in parallel in a culture apparatus.

  The microorganism culturing method according to claim 12, wherein the microorganism is formed on the upper surface of a porous body having a pore that is configured to allow liquid to pass therethrough and communicated with the surface, and wherein the upper surface pore is formed with a smaller pore diameter than the microorganism. A medium containing the sample is placed, the medium is continuously supplied to the porous body, the medium is passed in the direction intersecting the height direction inside the porous body, and the upper surface of the porous body is inside the porous body. A culture medium is supplied via the pores of, and microorganisms are cultured on the upper surface of the porous body.

  A microorganism culturing method according to claim 13 is the microorganism culturing method according to claim 12, wherein one solution selected from a plurality of solutions in which a plurality of the porous bodies are provided and different concentrations or different seeding sources are diluted. Is smeared on the upper surface of one porous body, a medium is supplied to each of the porous bodies smeared with the solution, and a plurality of cultures are performed in parallel.

  According to the microorganism culture device of the first aspect, the liquid is supplied to and discharged from the culture chamber through the flow path. With respect to the porous body held by the holding unit in the culture chamber, the liquid level is adjusted by the adjusting means so as to be the same height as the upper surface of the porous body or near the upper surface of the porous body and below the upper surface of the porous body, Direct liquid inflow from the outside to the upper surface of the porous body is restricted. On the other hand, since the porous body is configured to allow liquid to pass through, the liquid can flow through the porous body, and the supplied liquid passes through the pores inside the porous body due to the pores communicating with the surface. To the upper surface of the porous body.

  For this reason, the microorganisms seeded on the upper surface of the porous body can stay and grow at the position, and can avoid mixing with other microorganism cells. Further, since the porous body used has a pore diameter on the upper surface smaller than that of the microorganism, the microorganism growing on the upper surface of the porous body is not lost by the liquid flowing in the porous body. Furthermore, since the liquid can be continuously or intermittently supplied to the porous body, the nutrients necessary for the growth of the microorganism are not depleted, and the metabolite produced by the microorganism is continuously removed from the porous body. Since it can discharge | emit, the maintenance of the favorable microbial growth environment which was difficult with the conventional solid culture medium is realizable.

  Therefore, when culturing microorganisms with this device, it is possible to perform separation culture in which colonies of growing microorganisms are separated as single microbial cells in an environment where growth inhibition due to metabolites and depletion of nutrients are eliminated. it can. Furthermore, since the medium can be selected and changed during the culture, the degree of freedom in selecting the culture conditions is greatly improved and the culture conditions for various microorganisms are expanded compared to the conventional culture on a solid medium. There is an effect that can be made.

  In addition, the positional relationship between the position of the upper surface of the porous body and the level of the liquid to be supplied can be adjusted by the adjusting means. It can be arranged at the same height or near the upper surface of the porous body and slightly below the upper surface of the porous body.

  According to the microorganism culture device of claim 2, in addition to the effect produced by the microorganism culture device of claim 1, the pedestal fixed to the support column by the fixture is detachably attached to the support column. On the top, the porous body is detachably locked by the locking tool. Therefore, there is an effect that the porous body can be reliably fixed in the culture chamber into which the liquid flows. In addition, since the pedestal is detachably attached to the column, the column can be exchanged, and the height of the pedestal is changed by using a column having a different length, that is, the positional relationship between the upper surface of the porous body and the liquid level. Can be adjusted. For this reason, the positional relationship between the upper surface of the porous body and the liquid surface can be kept at a constant interval, and the liquid flowing into the culture chamber can be accurately avoided from flowing beyond the upper surface of the porous body.

  Furthermore, since the positional relationship between the upper surface of the porous body and the liquid surface can be adjusted by changing the structure of the pedestal, maintenance is facilitated by eliminating the need for an electric mechanism such as a motor, and the cost of the apparatus is further reduced. Can do.

  According to the microorganism culturing apparatus of claim 3, in addition to the effect produced by the microorganism culturing apparatus of claim 1, the pedestal attached to the support is fixed to the support so as to be movable in the vertical direction by a fixture, A porous body is detachably locked on the pedestal by a locking tool. The positional relationship between the upper surface of the porous body and the liquid level is adjusted by moving the pedestal in the vertical direction. Therefore, the porous body does not float unstablely in the culture chamber into which the liquid flows, and the porous body can be reliably fixed. That is, there is an effect that the positional relationship between the upper surface of the porous body and the liquid surface can be maintained at a constant interval, and the liquid flowing into the culture chamber can be reliably prevented from flowing beyond the upper surface of the porous body.

  Furthermore, by adjusting the fixture, the pedestal can be moved in the height direction to adjust the positional relationship between the upper surface of the porous body and the liquid surface, thus eliminating the need for an electric mechanism such as a motor and facilitating maintenance. The device cost can be reduced. Moreover, in this apparatus, the base can be continuously slid with respect to the support | pillar, for example, and there exists an effect that the operation | work can be simplified when performing fine adjustment of height.

  According to the microorganism culturing apparatus according to claim 4, in the microorganism culturing apparatus according to claim 2 or 3, the pedestal is provided with a through hole through which the liquid supplied to the culturing chamber passes in the thickness direction of the pedestal. Therefore, there is an effect that the supply of the liquid from the lower surface side toward the porous body can be prevented from being blocked by the pedestal. For this reason, in addition to the inflow of the liquid from the porous body side surface direction, the liquid can be supplied also from the lower surface side, so that the liquid can be efficiently supplied to the porous body.

  According to the microorganism culturing apparatus of claim 5, in addition to the effect exhibited by the microorganism culturing apparatus according to any one of claims 1 to 4, the holding body indirectly receives the porous body in the storage container. In addition to being able to hold, there is an effect that the liquid supply to the porous body can be satisfactorily performed through the communication portion that communicates the inside and outside of the container.

  Here, since the porous body is a structure having pores formed therein, it is more fragile than a dense structure. Moreover, since it is exposed to the flowing liquid, when it is directly gripped by the holding portion, stress concentration tends to occur in the gripped portion, and it may be damaged in some cases. In this apparatus, since the porous body is stored in the culture chamber after being accommodated in the storage container, even if the porous body is formed of a relatively weak material or a material that is difficult to directly grip, There is an effect that it can be held stably. Therefore, various porous bodies can be selected, and there is an effect that long-term culture can be realized as compared with the case of directly holding the porous bodies.

  According to the microorganism culturing apparatus of claim 6, in addition to the effects exhibited by the microorganism culturing apparatus according to any of claims 1 to 5, the porous body disposed in the culture chamber of the apparatus has a gel upper surface portion. Therefore, when a sample containing microorganisms is smeared on the upper surface of the porous body, which is a culture site, it can be smeared on a gel with good smoothness. In addition to being able to perform smoothly, there is an effect that liquid pool is less likely to occur and uneven coloring can be suppressed.

  According to the microorganism culturing apparatus of claim 7, in addition to the effects exhibited by the microorganism culturing apparatus of claim 6, since the porous body is formed entirely of gel, for example, gel and gel There is an effect that the production of the porous body can be made simpler and easier than the case where the porous body is formed by combining other materials than the above.

  According to the microorganism culturing apparatus according to claim 8, in addition to the effects exhibited by the microorganism culturing apparatus according to any one of claims 1 to 7, a bottomed housing that opens upward and a lid that closes the opening The culture chamber is contained in the outer tank equipped with the above, and the flow path is directly connected to the contained culture tank through the wall of the outer tank. That is, liquid can be supplied and discharged through the flow path to the culture chamber in the outer tank isolated from the outside. Therefore, there is an effect that the liquid can be supplied limitedly to the culture chamber in the outer tank.

  Therefore, in this apparatus, a place that does not come into contact with liquid can be secured in the internal space of the outer tank (the space between the inner wall of the housing and the culture chamber). For this reason, there exists an effect that the thing which should avoid water wetting, such as the substance which reacts with water, can be installed in this place in an outer tank, for example.

  In addition, since the opening of the outer tub can be switched between the open state and the closed state by the lid body, both aerobic and anaerobic environments can be realized, and the device is operated according to the atmosphere desired by the user. There is an effect that can be done.

  According to the microorganism culturing apparatus of claim 9, in addition to the effect exhibited by the microorganism culturing apparatus of claim 8, a sealing material that suppresses intrusion of outside air is provided at the opening portion of the casing of the outer tub. In addition, since the outer tub is configured so as to be able to be sealed when the lid is fixed by a jig, contamination from the outside such as contamination of germs can be suppressed. Furthermore, there is an effect that a favorable anaerobic atmosphere can be realized, and microorganisms that cannot grow unless the anaerobic degree is high can be favorably isolated and cultured.

  According to the microorganism culturing apparatus of the tenth aspect, in addition to the effect exhibited by the microorganism culturing apparatus according to the ninth aspect, the inside of the outer tank is made anaerobic by the oxygen scavenger provided in the inner space of the outer tank. be able to. Therefore, there is an effect that the inside of the outer tank can be made an anaerobic atmosphere in a simple and inexpensive manner as compared with the case where the replacement with nitrogen gas or the like is performed.

  According to the microorganism culture system of claim 11, a plurality of the microorganism culture devices according to any one of claims 1 to 10 are provided, and a flow path connected to each of the microorganism culture devices has The liquid is supplied from the storage tank by the supply pump, and the liquid is discharged from each of the microorganism culture apparatuses by the discharge pump. Thereby, there exists an effect that it can culture in parallel in each microorganism culture device. In addition, since the supply and discharge of liquid to and from each microorganism culture apparatus are performed by pumps, the flow rate can be easily adjusted and the desired flow rate can be adjusted compared to the case of supplying and discharging liquid by the action of gravity such as natural gradient. It is possible to achieve stable liquid (medium) supply and discharge in Furthermore, since the supply amount and discharge amount of the liquid can be controlled separately, there is an effect that the control and management of the liquid level in the culture chamber can be performed with high accuracy.

  According to the microorganism culturing method of claim 12, the upper surface of the porous body is configured so as to be permeable to liquid and having pores communicating with the surface, and the pores of the upper surface portion are formed with a pore diameter smaller than that of the microorganism. A sample containing microorganisms is placed on the porous body, the medium is continuously supplied to the porous body, and the medium is passed in the direction intersecting the height direction inside the porous body. The culture medium is supplied via the pores of, and the microorganism is cultured on the upper surface of the porous body. Therefore, since the culture medium (liquid medium) can be continuously supplied from the outside to the porous body serving as a culture site for microorganisms, there is an effect that nutrient sources necessary for the growth of microorganisms are not depleted. Moreover, compared with the case where it culture | cultivates with the solid medium like the former, the freedom degree with respect to selection and a change of a culture medium is large, and it can culture | cultivate on the conditions close | similar to the environment where microorganisms actually live.

  In addition, since the culture medium is supplied to the upper surface of the porous body via the pores inside the porous body, it does not flow on the upper surface of the porous body for culturing microorganisms. Therefore, disappearance of the formed colonies of microorganisms can be avoided. In addition, since microorganisms are cultured on the upper surface of the porous body and liquid supply such as a medium is supplied from the inside of the porous body to the upper surface of the porous body, it is possible to avoid a situation where microorganisms that grow with the supply of the medium are mixed. There is an effect that the microorganisms can be well separated.

  According to the microorganism culturing method of claim 13, in addition to the effects exhibited by the microorganism culturing method of claim 12, a plurality of porous bodies are provided and selected from a plurality of solutions in which different concentrations or different seeding sources are diluted. Since the above-mentioned 1 solution is smeared on the upper surface of one porous body, a medium is supplied to each of the porous bodies smeared with the solution, and a plurality of cultures are performed in parallel. Alternatively, there is an effect that the culture using a different seeding source can be efficiently performed.

It is a figure explaining the basic principle of the microorganism culture method of this invention. It is the figure which showed the outline | summary of the microorganism culture apparatus in 1st Embodiment of this invention. It is the figure which showed the outline | summary of the culture system containing the microorganisms culture apparatus of 2nd Embodiment of this invention. It is a figure explaining the culture tank of the microorganism culture apparatus in 2nd Embodiment. It is the figure which showed the storage container used by 2nd Embodiment. It is a figure explaining the height adjustment mechanism in the culture chamber in 2nd Embodiment. It is the figure which showed the modification of the mounting base of 2nd Embodiment. It is the figure which showed the microorganisms culture apparatus in 3rd Embodiment. It is the figure which showed the outline | summary of the microorganism culture system which is one Embodiment of this invention.

  DESCRIPTION OF EXEMPLARY EMBODIMENTS Hereinafter, preferred embodiments of the invention will be described with reference to the accompanying drawings. First, the basic principle of the microorganism culturing method of the present invention will be described with reference to FIG.

  FIG. 1 is a diagram for explaining the basic principle of the microorganism culturing method of the present invention. In this method, as shown in FIG. 1 (a), a porous body 1 is used as a microorganism carrier, and a microorganism group collected from the environment is appropriately diluted and smeared on the surface (upper surface) of the porous body 1. The microorganism is cultured above. Since the porous body 1 can permeate a liquid, a substrate serving as a nutrient source for microorganisms can be continuously supplied from the outside.

  The porous body 1 having liquid permeability is disposed in a liquid flow path. Liquid is introduced into the porous body 1 from the upstream side, and the liquid that has passed through the porous body 1 is discharged downstream. In addition, since supply and discharge | emission of a liquid should just be performed appropriately with respect to the porous body 1, supply of a liquid is not limited from the side of the porous body 1, For example, from the lower surface of the porous body 1. It may be done.

  FIG. 1B is a partially enlarged view I in the vicinity of the surface of the porous body 1. In the present invention, as shown in FIG. 1 (b), the liquid level is adjusted so as to be formed in the vicinity of the surface of the porous body (the same height h1 or less as the surface of the porous body 1 and above h2). Yes.

  If the liquid level exceeds the surface of the porous body 1, microorganisms seeded on the surface of the porous body may flow or may be mixed with other colonies, which may hinder acquisition of a single colony. For this reason, in this invention, it controls so that the upper limit of a liquid level may become a porous body surface.

  On the other hand, the porous body 1 includes pores through which liquid can permeate and micropores communicating from the inside to the surface. For this reason, even if the position of the liquid level is below the surface of the porous body, the liquid can be transported inside the porous body 1 and reach the surface by capillary action. However, if the liquid level is too lower than the surface position of the porous body 1, the supplied liquid does not reach the surface, and it becomes difficult to supply the liquid, that is, the nutrient source to the surface of the porous body. In addition, the efficiency of material exchange between the surface of the porous body and the inside of the porous body may decrease, and it becomes difficult to sufficiently remove the product produced by the microorganism from the surface of the porous body. For this reason, the limit point h2 (that is, the limit height at which the liquid can be transported upward by capillary action) where the material exchange with the surface of the porous body is favorably performed by the inflowing liquid is the lower limit position of the liquid surface. The limit point h2 is appropriately set according to the diameter of the micropores of the porous body 1, the amount of the supplied liquid, the flow rate, the viscosity, and the like, and is set, for example, 1 mm to several mm below the surface of the porous body 1.

  In addition, the micropores of the porous body 1 are preferably smaller than the size of the microorganism, and considering the efficiency of water flow, the pore diameter is preferably 0.05 μm to 10 μm, and more preferably Is from 0.1 μm to 1 μm. Since many microorganisms are larger than 1 μm, the use of a porous body having fine pores of 1 μm or less can prevent most microorganisms from falling from the surface of the porous body to the inside of the porous body. It is also possible to prevent the microorganism from advancing to the porous body surface when the microorganism propagates.

  The fine pores distributed in the porous body 1 do not necessarily have a uniform pore distribution over the entire porous body. The surface layer (portion having a predetermined thickness from the outermost surface) of the porous body 1 may be a layer having fine pores of 1 μm or less, and the lower layer may have pores larger than the fine pores of the surface layer. Thereby, it is possible to prevent microorganisms from moving from the surface layer of the porous body 1 (part having a predetermined thickness from the outermost surface) to the inside of the porous body, to cultivate microorganisms limited to the surface of the porous body 1, and to pass water. Efficiency can be improved.

  The porous body 1 is not particularly limited as long as it has fine pores and water permeability, and various organic polymer porous bodies and inorganic compound porous bodies can be used. Examples of the organic polymer porous body 1 include natural sponges such as sponges, artificial sponges made of urethane, melamine, rubber and the like, foams formed using organic polymers, non-woven fabrics, polymer films, Examples thereof include polymer gels in which molecules are crosslinked to form a network structure. Examples of the foam include, but are not limited to, molded materials using polyurethane, polystyrene, polyolefin, phenol resin, polyvinyl chloride, urea resin, silicone, polyimide, melamine resin, etc. as main raw material synthetic resins. It is not something that can be done.

  Examples of the nonwoven fabric include those formed using fibers such as aramid fiber, cellulose fiber, nylon fiber, vinylon fiber, polyester fiber, polyolefin fiber, and rayon fiber. Examples of the polymer film include cellulose acetate, three Cellulose acetate, cellulose nitrate, cellulose, polyacrylonitrile, polyamide, aromatic polyamide, polysulfone, polyethersulfone, polycarbonate, polyethylene terephthalate, polyimide, polyethylene, polypropylene, polytetrafluoroethylene, polyvinylidene fluoride, polyvinyl chloride, polyvinyl alcohol, etc. Although what is manufactured from can be illustrated, it is not restricted to this.

  Examples of polymer membranes include polyethylene resins, polypropylene resins, polyvinyl chloride resins, polyvinylidene fluoride resins, polysulfone resins, polyether sulfone resins, polyacrylonitrile resins, cellulose resins, and cellulose triacetate resins. Although what is formed using resin etc. is illustrated, it is not restricted to this.

  Furthermore, hydrogels are preferably used as the polymer gels, such as gels formed from polysaccharides such as agarose, cellulose, carrageenan, locust bean gum, guar gum, gellan gum, collagen gels, and polyvinyl alcohol. And synthetic polymers such as polyacrylamide, polyacrylamide, polyvinylpyrrolidone, polyhydroxymethyl acrylate, polyethylene oxide, acrylic acid, and methacrylic acid, and gels obtained by crosslinking these copolymers are exemplified. However, it is not limited to this. Moreover, these may use one type and may mix and use two or more types.

  In addition, it may be formed of an inorganic compound such as a ceramic porous body or a metal porous body. Examples of the ceramic porous body include a mesoporous body (porous glass), silica gel, silica alumina porous body, and activated carbon. Alumina diaphragm, ceramic filter, lightweight aggregate and the like can be exemplified, and examples of the metal porous body include those formed of nickel, titanium, cobalt, silver, stainless steel and the like, but are not limited thereto.

  The porous body 1 may be formed of a single layer or a plurality of layers, and each layer may be formed of the same material or different materials.

  Preferably, the surface layer portion of the porous body 1 is preferably formed of a gel. This is because the gel has good surface smoothness, so that the smearing operation of the liquid containing the microorganisms to be cultured can be performed smoothly, the liquid pool is hardly generated, and the unevenness of wetness can be suppressed. As a result, a liquid containing microorganisms can be thinly spread and formation of a single colony of microorganisms can be facilitated.

  In addition, if the porous body 1 is a gel, it is easy to form micropores of 1 μm or less, and it is a similar culture operation for users who have been cultured by the conventional agar plate surface smearing method. Is good. In addition, as for the porous body 1, only the surface layer part may be formed with the gel, and the whole porous body may be formed with the gel.

  In the present invention, the porous body 1 can be appropriately selected according to the culture conditions. For example, agar media that have been widely used so far cannot be cultured in high-temperature environments or environments with high acidity. However, in the present invention, for example, as a porous material having high heat resistance and high acid resistance, in addition to cellulose gel, synthetic polymer gel, and silica gel, mesoporous material (porous glass), silica alumina porous material, sponge Artificial sponges such as urethane foam and melamine foam, non-woven fabrics of aramid fibers, cellulose fibers, and nylon fibers can be used. In particular, silica gel is a porous gel plate that can be produced in a range of about pH 5 to 9.5, and can be cultured in an acidic or alkaline extreme environment.

  As a result, it is possible to artificially reproduce the culture environment that mimics the extreme environment and perform separation culture of microorganisms, and it is possible to realize separation culture of microorganisms in the extreme environment and non-separable microorganisms, which has been difficult until now .

  As described above, if the microorganism culture method of the present invention is used, microorganisms can be cultured in the same manner as a solid medium while continuously supplying a substrate (culture solution). It is possible to avoid that microorganisms cannot grow due to depletion of, and that growth inhibition occurs due to accumulation of microbial product. In addition, unlike the case where microorganisms are cultured inside a porous material such as perfusion culture, liquid does not flow on the surface of the porous material where microorganisms are cultured. Can be earned. Furthermore, the liquid (culture solution composition) can be changed and adjusted, and the culture and acclimatization of microorganisms can be performed by reproducing the medium composition and culture conditions that more closely approximate the natural environment. In addition, the culture environment can be diversified, and separation and culture can be performed under a wider range of conditions than in the conventional method.

  Next, with reference to FIG. 2, the microorganism culture apparatus in 1st Embodiment of this invention is demonstrated. In addition, the following embodiment is only an example which actualized this invention, and it cannot be overemphasized that embodiment can be changed suitably in the range which does not change the summary of this invention.

  The microorganism culturing apparatus 3 is for holding the above-mentioned porous body 1 inside and performing microorganisms separation culture on the surface of the porous body. The microorganism culture apparatus 3 includes a culture chamber 3a in which the porous body 1 is accommodated. The porous body 1 is accommodated (held) in an accommodation container X having a plurality of through holes on at least a side surface and has a predetermined height. Stored in the culture chamber 3a. In FIG. 2, the storage container X is placed on a base having a predetermined height and is held at a predetermined height. However, the present invention is not limited to this, and the storage container is placed in the culture chamber 3a using a hanging metal fitting. X may be suspended, and various methods of fixing a plurality of coil springs on the side surface of the storage container X and fixing the storage container X to the inner wall of the culture chamber 3a by the biasing force can be applied.

  In addition, the petri dish mentioned later in 2nd Embodiment may be used for the storage container X, and the mounting base 20 mentioned later in 2nd Embodiment may be used as a holding means to hold | maintain a storage container.

  The culture chamber 3a is a glass container provided with a bottomed cylindrical main body having an open upper surface and two through holes formed in the peripheral wall of the main body. The culture chamber 3a forms a part of the flow path through which the liquid flows, and the container X for the porous body 1 is installed as described above, and serves as a chamber for culturing microorganisms using the porous body 1.

  The two through holes are formed facing each other in the diameter direction of the cylindrical main body. One through hole is disposed on the upstream side of the flow path and is connected to the supply unit 3b. The other through hole is disposed on the downstream side of the flow path and is connected to the discharge portion 3c. The supply unit 3b and the discharge unit 3c are hollow conduits protruding outward from the peripheral wall of the main body of the culture chamber 3a, and are formed integrally with the main body of the culture chamber. The supply unit 3b communicates with the through hole in the hollow portion inside, and introduces the culture solution from the outside to the culture chamber 3a through the through hole. The discharge part 3c communicates with the through hole in the hollow part inside, and discharges the culture solution from the culture chamber 3a to the outside through the through hole.

  Here, in this embodiment, the supply part 3b is provided in the position lower than the discharge part 3c. Thereby, the water level of the culture solution supplied from the supply part 3b rises to the position of the discharge part 3c, and is discharged out of the culture chamber from the position of the discharge part 3c. As a result, the culture solution can be supplied to a position near the surface of the porous body 1 and slightly lower, and a liquid surface with a certain height is formed with respect to the surface of the porous body 1 in the culture chamber 3a. And it is possible to avoid the occurrence of an extreme height gradient on the liquid surface of the culture solution. In addition, when the liquid level of fixed height can be formed with respect to the surface of the porous body 1, the supply part 3b does not necessarily need to be provided in a position lower than the discharge part 3c. You may provide 3b and the discharge part 3c at the same height.

  If the microorganism culture apparatus 3 is installed in the flow of the culture solution and fixed so that the liquid level in the culture chamber is held at a position slightly lower than the upper surface of the porous body 1, the substrate is continuously porous. The body 1 can be supplied. That is, the microorganism can be cultured on the surface of the porous body while continuously supplying the substrate. Alternatively, a tube may be attached to the tip of the supply unit 3b and the discharge unit 3c, and the culture solution (substrate) may be transported to the culture chamber 3a through the tube by driving the pump.

  In addition, the culture solution to be supplied is not necessarily limited to one in which a nutrient source is artificially blended and adjusted, and may be an aqueous liquid in an actual environment such as seawater, a river, or treated water of another plant. In the case of using such an aqueous liquid in the actual environment, it is desirable that a sterilization treatment such as an autoclave treatment is performed in order to avoid contamination.

  Next, with reference to FIG. 3 to FIG. 6, a microorganism culturing apparatus in the second embodiment of the present invention will be described.

  FIG. 3 is a schematic diagram showing an outline of the culture system 10 including the microorganism culture device 4 in the second embodiment. The culture system 10 is configured to be able to separate and culture microorganisms while continuously supplying a culture solution (liquid) to the microorganism culture apparatus 4, and in order from the upstream side, a storage tank 11, a supply pump 12, and a microorganism culture apparatus. 4, a discharge pump 14 and a drainage tank 15 are provided. Further, a delivery pipe 16 for delivering the culture solution from the storage tank 11 to the drainage tank 15 is provided, and this delivery pipe 16 is a supply tube 16a for supplying the culture solution from the storage tank 11 to the microorganism culture apparatus 4. And a discharge tube 16 b that discharges the culture solution from the microorganism culture device 4 to the drainage tank 15.

  The storage tank 11 stores a culture solution to be supplied to the culture tank 13, and is formed of a screw mouth bottle provided with a screw mouth-shaped wide-mouth glass bottle main body 11a and an upper lid 11b screwed to the screw mouth. Yes. A through hole into which a hollow suction tube 11c is fitted is formed in the upper lid 11b, and the suction tube 11c is inserted into the wide-mouth glass bottle main body 11a through the through hole. The suction tube 11c has a lower end disposed in the vicinity of the bottom surface in the wide-mouth glass bottle body 11a, and an upper end formed with a predetermined length so as to protrude outward from the upper lid 11b, and is fitted into the upper lid 11b. The culture solution stored in the wide-mouthed glass bottle body 11a is led out to the outside through the suction tube 11c. For this reason, one end of the supply tube 16a is connected to the upper part of the suction pipe 11c. In addition, when supplying seawater, a river, the treated water of another plant, etc. to the culture tank 13 as a culture solution, this liquid is stored in this storage tank 11 after performing sterilization processes, such as an autoclave process. .

  The wide-mouth glass bottle main body 11a, the upper lid 11b, and the suction pipe 11c are each made of a sterilizable material, and the wide-mouth glass bottle main body 11a and the suction pipe 11c are made of borosilicate glass. The upper lid 11b is made of a polypropylene resin that can be sterilized by an autoclave. There is no particular limitation as long as it can be used in a sterilized state, and the wide-mouth glass bottle main body 11a and the suction tube 11c may be made of resin. Examples of the resin that can be sterilized by autoclave include, in addition to the above-described polypropylene resin, phenol resin, fluorine resin, polycarbonate resin, polymethylpentene resin, and the like. Further, not only autoclave sterilization but also sterilization by electron beam irradiation may be used. In such a case, a general-purpose resin having a relatively low heat resistance such as a polyethylene resin can be used.

  In the main culture system 10, the wide-mouth glass bottle body 11a, the upper lid 11b, and the suction tube 11c are sterilized by autoclaving, and then the sterilized culture solution is introduced into the wide-mouth glass bottle body 11a, and the upper lid 11b After the fitting, the suction tube 11c is attached. Aseptic conditions are maintained by performing a series of operations aseptically.

  The supply tube 16 a is a flow path for supplying the culture solution stored in the storage tank 11 to the culture tank 13. One end of the supply tube 16 a is connected to the suction pipe 11 c of the storage tank 11 and extends through the supply pump 12. It is a resin tube that connects the other end to the microorganism culture apparatus 4. The supply tube 16a is a general one that can be sterilized. For example, the supply tube 16a is a resin tube made of a material such as polyvinyl chloride resin, silicon rubber, fluorine rubber, styrene-ethylene-butylene elastomer, or polypropylene elastomer. Therefore, those suitable for the culture medium are appropriately selected and used. The supply tube 16a is sterilized by autoclaving or the like, and then aseptically operated, one end thereof is connected to the suction tube 11c and the other end is connected to the microorganism culture apparatus 4.

  The supply pump 12 is a tubing pump that can control the flow rate. Although not shown, the supply pump 12 is provided with a power switch for switching the power on and off, a dial for changing the rotation speed (that is, the flow rate), and a reversing switch for reversing the flow direction. ing. And it is a general apparatus provided with the pump head in which the supply tube 16a is set.

  The pump head includes a substantially disk-shaped rotor having a plurality of rollers at the outer edge, and the center of the rotor is pivotally attached to the rotation shaft of the drive unit. The pump head is formed with a groove for installing the supply tube 16a so as to contact the roller. When the power is turned on, the rotor rotates and the roller moves while crushing the supply tube 16a. The culture solution in the tube is pushed out in the moving direction. Thereby, a culture solution is sent out downstream. The supply pump 12 is configured to be connected with a plurality of pump heads, and can supply a culture solution to a plurality of delivery destinations.

  The microorganism culturing apparatus 4 has a substantially cylindrical outer shape, and is a culture chamber 13 that is a glass chamber formed in an openable and closable manner that is sealed by a fixture 40, and a culture provided in the culture tub 13. Chamber 30.

  The culture chamber 30 includes a bottomed cylindrical main body having an open upper surface, a supply unit 31 and a discharge unit 32 projecting outward from the peripheral wall of the main unit. It is a compartment made of glass. The culture chamber 30 forms a part of a flow path through which a liquid flows, and a petri dish 2 that is a container for the porous body 1 is installed, and serves as a chamber for culturing microorganisms using the porous body 1.

  The supply unit 31 is a hollow conduit for introducing the culture solution from the outside of the culture vessel 13 into the culture chamber 30, and the discharge unit 32 is a hollow for introducing the culture solution from the culture chamber 30 to the outside of the culture vessel 13. This is a conduit. The supply part 31 and the discharge part 32 are arrange | positioned facing the diameter direction of a cylindrical main body. The bases of the supply unit 31 and the discharge unit 32 are fused to the peripheral wall of the culture chamber 30 main body. In the fused portion of the supply portion 31 and the discharge portion 32 on the peripheral wall of the main body, a drill hole having a diameter substantially the same as the inner diameter of the supply portion 31 and the discharge portion 32 is formed. For this reason, the flow paths 31 a and 32 a (see FIG. 4) formed inside the supply unit 31 and the discharge unit 32 respectively communicate with the culture chamber 30. As in the first embodiment, the supply unit 31 and the discharge unit 32 are provided at a position where the supply unit 31 is lower than the discharge unit 32, and thereby, the culture solution at a constant height in the culture chamber 30. The liquid level is formed.

  Further, the tips of the supply unit 31 and the discharge unit 32 extend through the side wall of the culture tank 13 to the outside, and the other end of the supply tube 16 a is connected to the tip of the supply unit 31. . Thus, when the supply pump 12 is driven, the culture solution is introduced from the supply unit 31 to the culture chamber 30 in the culture tank 13 via the supply tube 16a.

  Furthermore, one end of the discharge tube 16 b is attached to the tip of the discharge portion 32. The discharge tube 16b is a flow path for sending the culture solution discharged from the culture tank 13 to the drain tank 15, and is configured in the same manner as the supply tube 16a. The discharge tube 16 b is connected to the drainage tank 15 via the discharge pump 14.

  The discharge pump 14 is a pump for discharging the culture solution from the culture tank 13 (that is, the culture chamber 30), and is configured similarly to the supply pump 12. By providing the discharge pump 14, it is possible to stably supply the culture solution in a fixed amount as compared with the case where only the supply pump 12 is provided. That is, in the main culture system 10, the culture medium supplied to the culture chamber 30 can be strictly controlled by providing the supply pump 12 and the discharge pump 14. The surface height can be precisely controlled. In the culture system 10, the storage tank 11 is provided at a position higher than the culture tank 13, the drainage tank 15 is disposed at a position lower than the culture tank 13, and the culture solution is supplied to the culture tank 13 due to the height difference. If the height of the culture liquid in the culture chamber 30 can be adjusted to a specified range by such a configuration, at least one of the discharge pump 14 or the supply pump 12 is not provided. It is good. In the main culture system 10, the discharge rate of the culture solution by the discharge pump 14 is set slightly higher than the supply rate of the culture solution by the supply pump 12. Thereby, it can be strictly controlled that the liquid level of the culture solution formed in the culture chamber 30 exceeds the surface of the porous body 1.

  The supply rate of the culture solution can be appropriately selected according to the type of microorganisms intended for culture and the concentration and viscosity of the culture solution.

  The drainage tank 15 collects the culture solution discharged from the culture tank 13, and is formed of a screw mouth bottle provided with a screw mouth-shaped wide-mouth glass bottle main body 15a and an upper lid 15b screwed to the screw mouth. Has been. A through hole into which a hollow discharge pipe 15c is fitted is formed in the upper lid 15b, and the discharge pipe 15c is inserted into the wide-mouth glass bottle main body 15a through the through hole. The discharge pipe 15c has a lower end disposed in the wide-mouth glass bottle main body 15a and is formed with a predetermined length so that the upper end protrudes outward from the upper lid 15b, and is fitted to the upper lid 15b. It is a cylindrical tube, and the other end of the discharge tube 16b is connected to the upper end portion. Thereby, the culture solution sent from the culture tank 13 via the discharge tube 16b flows through the discharge tube 15c and is stored in the wide-mouth glass bottle body 15a.

  Each part of the drainage tank 15 is formed of a sterilizable material similar to that of the storage tank 2, and in the main culture system 10, the wide-mouth glass bottle body 15a, the upper lid 15b, and the discharge pipe 15c are sterilized by an autoclave. After being performed, the exhaust pipe 15c is attached after being tightly fitted by the upper lid 15b. Aseptic conditions are maintained by performing a series of operations aseptically.

  Here, in the culture system 10, the substance produced by the microorganism is discharged from the culture tank 13 together with the culture solution and stored in the discharge tank 15, so that a desired product substance is obtained from the culture solution stored in the discharge tank 15. It can be recovered. Note that the drainage tank 15 may be omitted when the drainage is discharged out of the system as it is. Furthermore, the culture solution may be circulated by disposing the drainage tank 15 and connecting the discharge tube 16b to the storage tank 11.

  Next, with reference to FIG. 4, the detailed structure of the culture tank 13 is demonstrated.

  FIG. 4 is a diagram illustrating the configuration of the culture tank 13, and FIG. 4A is a cross-sectional view of the culture tank 13. As shown to Fig.4 (a), the culture tank 13 is provided with the storage tank 13a for accommodating said culture chamber 30, and the cover body 13b which covers the upper surface of the storage tank 13a, and comprises the storage tank 13a. By covering with the lid 13b, the inside is separated from the outside.

  Specifically, the storage tank 13a is formed in a substantially bottomed cylindrical shape, and the culture chamber 30 is stored. As described above, the supply unit 31 and the discharge unit 32 are provided so as to pass through the side wall of the storage tank 13a (the side wall of the culture tank 13), respectively, so that the airtightness in the culture tank 13 is maintained. In the part, the supply part 31, the discharge part 32, and the side wall of the storage tank 13a are fused without a gap. Further, the supply section 31 and the discharge section 32 are fused to the storage tank 13a in this way, so that the culture chamber 30 is joined to the storage tank 13a and suspended in the storage tank 13a.

  In the opening on the upper side of the storage tank 13a, a peripheral edge having a smooth upper surface arranged flush with the opening surface and extending outward from the edge of the opening is provided around the opening of the storage tank 13a. ing. The peripheral edge is formed integrally with the storage tank 13a, and a groove 13c1 is formed in an annular shape around the peripheral edge.

  The lid 13b has a cylindrical casing having a top plate, and is used by placing the lower part of the casing on the upper surface of the storage tank 13a. The inner diameter and outer diameter of the housing are formed to be the same as the inner diameter and outer diameter of the storage tank 13a, respectively. In addition, the opening on the lower side of the lid body 13b has a peripheral portion that has a smooth surface that is flush with the opening surface and extends outward from the edge of the opening. It is installed. The peripheral edge is formed integrally with the lid 13b, and comes into contact with the peripheral edge of the storage tank 13a when placed on the storage tank 13a. In addition, a groove 13c2 is formed in a circular shape around the periphery of the lid 13b. The groove 13c2 is provided at a position facing the groove 13c1 on the storage tank 13a side when the lid 13b is placed in the storage tank 13a. Is set with an annular sealing rubber 13 d for improving the airtightness of the culture tank 13. For the sealing rubber 13d, a material having a low gas permeability is used, and for example, butyl rubber or fluorine rubber is preferably used. The lid 13b is detachably fixed to the storage tank 13a by the fixture 40 and seals the culture tank 13.

  FIG. 4B is a diagram illustrating the configuration of the culture chamber 30 provided in the culture tank 13. The culture chamber 30 is a chamber for holding a petri dish 2 containing the porous body 1 therein and culturing microorganisms with the porous body 1. For this reason, the petri dish cover 17 which covers the upper surface of the petri dish 2 and the mounting table 20 on which the petri dish 2 is placed are arranged together with the petri dish 2.

  That is, the petri dish 2 is detachably held in the culture chamber 30. For this reason, the porous body 1 can be installed in the culture chamber 30 with the petri dish 2 set in advance and can be exchanged. When the porous body 1 is formed of a gel or when a microbial sample is smeared on the surface of the porous body, a delicate operation is required for handling the porous body. In the present embodiment, since the porous body 1 can be taken in and out of the petri dish 2 in the state in which the porous body 1 is accommodated, the porous body 1 can be directly set (collected) in the culture chamber 30. Compared with the case where the porous body 1 is a gel or the case where a microbial sample is smeared on the surface, high caution and skill are not required and the operation can be easily performed.

  Here, the petri dish 2 which is a storage container used in the present embodiment will be described with reference to FIG. FIG. 5 is a diagram showing an outline of the petri dish 2. The petri dish 2 is a bottomed cylindrical thin-walled resin container having an open upper surface. A plurality of openings 2 a are formed on the side surface of the petri dish 2 erected from the bottom end. The main culture system 10 is configured to dispose the petri dish 2 in a liquid flow path and supply the liquid continuously or intermittently to the porous body 1 accommodated in the petri dish 2. Here, the liquid supplied from the upstream side, that is, the supply unit 31 (flow path 31a) flows into the porous body 1 in a state not exceeding the surface of the porous body 1 and permeates through the porous body 1 to One or more openings 2 a are provided on each of the upstream side and the downstream side so as to be discharged from the body 1.

  Furthermore, the petri dish 2 has a large through hole 2b on its bottom surface. Thereby, the liquid can be supplied to the porous body 1 from the lower surface side, and the liquid can be efficiently supplied to the porous body 1 accommodated in the petri dish 2.

  Returning to FIG.

  The petri dish cover 17 is a resin cover provided with a disk-shaped top plate having a diameter longer than the diameter of the upper surface of the petri dish 2 and a peripheral wall formed at a predetermined height downward from the top plate end. It is. The top plate of the petri dish cover 17 is formed with a through hole 17a into which a fixing screw 18 is inserted at a position outside the petri dish 2 in a state where the petri dish cover 17 is stacked on the petri dish 2.

  The mounting table 20 is used for mounting the petri dish 2 and is formed of a material that can be sterilized and made of a material having a specific gravity that does not float in the supplied liquid. As such a material, for example, resin, metal, ceramics, and glass can be used. The resin material preferably has a specific gravity of 1.2 or more, for example, polycarbonate resin, polysulfone resin, polyetherimide resin, polyurethane resin, polyethylene terephthalate resin, polybutylene terephthalate resin, fluorine resin. Resins and the like are exemplified. Moreover, you may use what added fillers, such as an inorganic substance, to the resin material, and improved specific gravity. In addition, a material resistant to autoclave sterilization (121 ° C., 2 atm) is preferably selected. In the present embodiment, the mounting table 20 is made of polytetrafluoroethylene, which is a fluorine-based resin. For this reason, the mounting table 20 is seated by its own weight, and the stationary state is maintained even if the culture solution is supplied into the culture chamber 30.

  The mounting table 20 includes an upper structure 21, leg portions 22, and countersunk screws 23. The upper structure 21 has a disk-shaped bottom surface 21a and a side surface 21b erected from the periphery of the bottom surface 21a, and is formed in a shallow basin shape. In addition, a relatively large through hole 21c is formed at the center of the bottom surface 21a to the extent that it does not interfere with the placement of the petri dish. Further, although not shown, the side surface 21b is notched. Is provided.

  The culture solution supplied from the supply unit 31 (channel 31a) into the culture chamber 30 needs to flow into the upper structure 21 of the mounting table 20. Although the height of the side surface 21b is lower than the height of the petri dish 2, a cutout is provided in the side surface 21b so that the liquid flows more efficiently into the upper structure 21. Furthermore, a through hole 21c is formed to create a liquid inflow path from the lower surface side.

  As a result, the liquid that has flowed into the upper structure 21 passes through the opening 2a and the through hole 2b of the petri dish 2 and is supplied to the inside of the porous body 1, and from the porous body 1 to the opening 2a and the through hole 2b. It is discharged smoothly via.

  In addition, the bottom surface 21 a of the upper structure 21 is provided with a drilling hole 21 d drilled in the thickness direction at a position where the leg portion 22 is disposed. The hole 21d is formed in a mortar shape whose inner diameter becomes narrower as it goes downward so that a countersunk screw 23 inserted to fix the leg 22 is locked. Further, the drill holes 21d are arranged on the outer peripheral side of the through holes 21c, and are provided at three positions so that the upper structure 21 is evenly supported by the three legs 22.

  Furthermore, when the petri dish 2 is placed on the bottom surface 21a, a screw is placed at a position that is outside the petri dish 2 and that faces the through hole 17a when the petri dish 2 is covered with the petri dish cover 17. A hole 21e is formed. Inside the screw hole 21e, a female screw into which the fixing screw 18 which is a male screw is screwed is screwed. The fixing screw 18 has a head having a diameter larger than that of the through hole 17a, and the shaft has a length that reaches (screws in) the screw hole 21e when inserted into the through hole 17a from the upper surface of the petri dish cover 17. Have. Two screw holes 21e and two through holes 17a are provided, and the petri dish cover 17 covers the petri dish 2 when the two fixing screws 18 are screwed into the screw holes 21e through the through holes 17a. Attached to state and fixed. Accordingly, the petri dish 2 is sandwiched between the bottom surface 21 a of the upper structure 21 and the petri dish cover 17 and is fixed (stablely held) in a fixed position in the culture chamber 30.

  Since formation of the opening 2a and the through-hole 2b is easy and it is cheaper, when the petri dish 2 is a resin product, it is easily levitated in the liquid because it is lightweight. When the petri dish 2 is in a floating state, the liquid surface position with respect to the porous body 1 is not stable, and it becomes difficult to control the liquid supply to the vicinity of the surface of the porous body 1. Therefore, in the main culture system 10, a structure that can fix the petri dish 2 is adopted, the surface position of the porous body 1 is positioned, and the positional relationship between the surface and the liquid level is made constant. Thereby, the liquid to be supplied can be accurately supplied to the porous body 1 so that the liquid to be supplied has a height not exceeding the surface of the porous body 1 and forms a liquid surface in the vicinity of the surface of the porous body 1. .

  The leg 22 is formed of a cylindrical body, and a screw hole 22a in which a female screw corresponding to the countersunk screw 23 is threaded is formed in the cylindrical body along the height direction. The leg portions 22 are arranged so as to face the respective drilling holes 21d of the upper structure 21, and the countersunk screws 23 are screwed into the screw holes 22a from the drilling holes 21d to be fastened. It is attached and fixed to the structure 21. As described above, since the mounting table 20 is made of polytetrafluoroethylene having a relatively large specific gravity, the mounting table 20 can be stably placed without floating even if disposed in the culture chamber 30 through which the liquid flows.

  In the present embodiment, as described above, the upper structure 21 is supported by the three legs 22. As a result, a large escape area for the liquid flowing into the culture chamber 30 is secured, and the force (resistance) received by the legs 22 (that is, the mounting table 20) from the liquid is reduced, so that the mounting table 20 in the culture chamber 30 can be reduced. Stability can be improved.

  FIG. 6 is a view for explaining a height adjusting mechanism in the culture chamber 30. In the present embodiment, the petri dish 2, that is, the surface position (height) of the porous body 1 is adjusted using the legs 22. Specifically, as shown in FIG. 6, the legs 22 are provided with a plurality of types having different lengths. Further, a plurality of types of countersunk screws 23 corresponding to each leg portion 22 are provided. Each countersunk screw 23 that is paired with each leg 22 has such a length that the screw tip does not protrude from the tip (lower end) of the leg 22 when the leg 22 is fastened to the upper structure 21. , Each is formed.

  The legs 22 are of the same type (length) as a set of three, and each leg 22 is attached to the upper structure 21 with a countersunk screw 23 that forms a pair. That is, since the leg part 22 can be attached to and detached from the upper structure 21 by the countersunk screw 23, the height of the leg part 22 can be adjusted stepwise by using the leg part 22 of a desired length as appropriate during use. Can do. In FIG. 6, three types of leg portions 22 are shown. However, more types (lengths) of the leg portions 22 may be provided. Further, the number of leg portions 22 attached to the upper structure 21 is not limited to three, and may be two or four or more. In such a case, the perforation holes 21 d of the upper structure 21 are also provided according to the number of leg portions 22.

  As described above, the height adjustment mechanism of the present embodiment realizes the positional relationship adjustment between the surface of the porous body 1 and the liquid level by adjusting the height of the mounting table 20. In other words, the height change of the mounting table 20 is configured to be performed only by mechanical adjustment without using an electric drive source, so that the height adjustment can be performed even in an environment exposed to liquid. The mechanism can be easily arranged, and the maintenance can be simplified.

  In the present embodiment, various types of porous body 1 can be used, and even if the same type is used, the porous body 1 always has the same thickness because it is affected by the difference in lots during manufacturing and the difference in production conditions. The body 1 cannot be used. In addition, it is also assumed that the reaching force (penetration force) from the inside of the porous body 1 to the surface varies depending on the flow rate and viscosity of the supplied liquid. In the present invention, it is important to form a liquid surface at a height not exceeding the surface of the porous body 1 and in the vicinity of the surface. For this reason, even if the thickness of the porous body 1 and the conditions of the culture solution change, a height adjusting mechanism is provided in order to make the positional relationship between the surface of the porous body 1 and the liquid level within an appropriate range. And in this embodiment, this height adjustment can be simply performed now by providing the leg part 22 from which length differs in the upper structure 21 so that attachment or detachment is possible.

  As described above, according to the microorganism culturing apparatus 4 of the present embodiment, the culture solution of the cultured microorganism can be configured to be continuously supplied to the porous body 1, and from the outside (according to the culture system 10) during the culture period. ) The supply amount and composition of the culture solution can be arbitrarily changed. In other words, in the agar plate surface smearing method used for normal separation culture, culturing must be performed in a predetermined defined medium, and thus the culturable microorganism species is limited. If used, the microbial species that can be cultured can be increased. Moreover, according to the microorganism culturing apparatus 4, since the cultured microorganisms are not community microorganisms and can be continuously cultured with individual colonies separated (single colonies are formed), useful microorganisms Can be separated individually.

  In the second embodiment, the height adjusting mechanism is provided in the culture chamber 30, and the height (positional relationship between the porous body 1 and the liquid surface) is changed by mechanical adjustment. The position of the body 1 may be a fixed position, and the liquid surface position may be changed by controlling the flow rate of the liquid introduced into the culture chamber 30. Alternatively, the height may be adjusted by an electric lifting mechanism. The elevating mechanism may be, for example, built in the support or may be exposed to the outside, and is introduced into the culture tank 13 after being sterilized as necessary.

  Furthermore, in the second embodiment, the entire system is configured as a closed system. However, the microorganism culture apparatus 3 is used instead of the microorganism culture apparatus 4, and the culture is performed in a state where the aerobic atmosphere is maintained. May be. In such a case, a sterilization filter may be placed over the opening of the culture chamber 3a in order to prevent invasion of various bacteria. Further, the entire apparatus may be placed in a clean bench for culturing.

  In addition, the inside of the microorganism culture apparatus 4 may be aerobic, microaerobic, or anaerobic, and may be in a specific gas environment. Such an in-apparatus environment can be realized by gas replacement. Examples of the gas to be replaced include carbon dioxide gas, nitrogen gas, sulfur gas, and methane gas. Various gases and mixed gases thereof can be used depending on microorganisms desired to be cultured. Can be appropriately selected.

  Moreover, in the microorganism culture apparatus 4 of the said 2nd Embodiment, you may add the structure which installs a light source and a sensor above the culture tank 13, and performs colony count of the microorganisms which appeared on the surface of the porous body 1. FIG. Thereby, for example, if the drug is mixed with the liquid supplied to the porous body 1 and the colonies of the growing microorganisms are counted, the drug resistance of the cultured microorganisms can be detected. When the culture tank 13 is transparent, the light source and the sensor may be provided outside the culture tank 13 or may be installed in the culture tank 13 after sterilization.

  Furthermore, in the said 2nd Embodiment, the height adjustment mechanism provided in the culture chamber 30 replaces | hangs the leg part 22 of the mounting base 20 with the thing from which length differs, and is supplied with the surface position of the porous body 1. The positional relationship with the liquid level of the liquid to be adjusted was adjusted. Instead of this, the height may be adjusted by a mounting table 200 as shown in FIG.

  FIG. 7 is a view showing the mounting table 200. The mounting table 200 is made of polytetrafluoroethylene and includes a disk-shaped upper structure 210, a nut 220, and a bolt 230. The upper structure 210 is provided with a screw hole 210 e similar to the screw hole 21 e provided in the mounting table 20, so that the petri dish cover 17 can be fixed with the fixing screw 18. A through hole 210 c similar to the through hole 21 c of the mounting table 20 is formed. Further, a hole 210 d through which the bolt 230 is inserted is formed through in the thickness direction of the upper structure 210.

  The upper structure 210 is supported using three bolts 230 as columns. Therefore, three holes 210d are provided in the upper structure 210. The holes 210d are located outside the petri dish 2 when the petri dish 2 is placed in the upper structure 210, and are arranged at substantially equal intervals on concentric circles.

  Each bolt 230 is formed with a sliding feed screw, and is fastened to the upper structure 210 using a pair of nuts 220. Since the position of the nut 220 attached to the bolt 230 can be moved by rotating, the position of the nut 220 attached to each bolt 230 is rotated to move the position, and then the other nut is moved. By following and moving, the holding position of the upper structure 210 can be changed and fixed (that is, the upper structure 210 is moved up and down). Thereby, the height of the petri dish 2 to be placed, that is, the positional relationship between the surface position of the porous body 1 and the liquid level can be adjusted.

  In this modification, the height adjustment mechanism is configured by the bolt 230 and the nut 220. However, any combination of the bolt 230 and the nut 220 may be used as long as the mechanism can move the upper structure 210 in the vertical direction. It is not limited. For example, the support column that supports the upper structure 220 is a cylindrical body having a plurality of holes drilled in the height direction (side surface) instead of the bolt 230, and a pin that is inserted into the hole of the cylindrical body instead of the nut 220. It is good also as a structure which supports the upper structure 220 using. Here, if the top of the pin has a length that extends outward from the hole, the upper structure 220 can be supported by the pin, and the position of the pin can be moved to move the upper structure 210. The height can be varied. Furthermore, it is good also as a structure which changes the height of the upper structure 210 by combining the clip and support | pillar which can be slid to an up-down direction, supporting the upper structure 210 with the said clip, and sliding a clip.

  Next, with reference to FIG. 8, the microorganism culture apparatus 100 of 3rd Embodiment is demonstrated. In addition, the same code | symbol is attached | subjected to the same part as the microorganism culture apparatus 4 of above-described 2nd Embodiment, and the description is abbreviate | omitted.

  FIG. 8 is a diagram illustrating the culture tank 13 of the microorganism culture device 100 according to the third embodiment. FIG. 8A is a view showing the appearance of the culture tank 13 of the microorganism culture apparatus 100, and FIG. 8B is a top view of the culture tank 13 of the microorganism culture apparatus 100. FIG. In the microorganism culture apparatus 100 of the second embodiment, the oxygen scavenger 50 is set in the culture tank 13. Thereby, the environment in a tank can be simply made anaerobic.

  Further, a material with low gas permeability is selected for the supply tube 16a in order to facilitate anaerobic retention, and butyl rubber or fluorine rubber is preferably used. As in the second embodiment, the supply tube 16a is sterilized by autoclaving or the like, and then one end is connected to the suction tube 11c and the other end is connected to the culture tank 13 by aseptic operation.

  The oxygen scavenger 50 is a bag body formed of a gas-permeable sheet body containing a drug that absorbs oxygen, and absorbs oxygen in the air with the drug through the sheet body. is there. In the present embodiment, a drug mainly containing ascorbic acid is used as a drug that absorbs oxygen. The oxygen scavenger 50 is not particularly limited as long as it has an action of absorbing oxygen, and a carbon dioxide gas generating agent and an oxygen absorbent that are used for general purposes can be used. The culture tank 13 has a good sealing property and a structure with excellent airtightness. For this reason, by setting the oxygen scavenger 50 inside the tank, the tank environment can be made anaerobic.

  For example, by introducing nitrogen gas or the like into the culture tank 13, the inside of the tank can be made an anaerobic environment. However, complicated operations are required to perform gas replacement, and skill is required for the work. Furthermore, since nitrogen gas generally used for gas replacement is expensive, the cost increases. Therefore, in this embodiment, an anaerobic environment can be easily realized by using an oxygen scavenger.

  Here, the culture tank 13 needs to have a structure in which a liquid flows. For this reason, in the culture tank 13, the culture chamber 30 through which the liquid flows is partitioned and configured so that the liquid does not flow outside the culture chamber 30 (outside the liquid flow path). ing. In addition, each size is designed so that a gap is formed between the outer wall of the culture chamber 30 and the inner wall of the culture tank 30, and the oxygen scavenger 50 is set in the gap. ing.

  Thereby, it is avoided that the oxygen scavenger 50 is immersed in the liquid flowing in the culture chamber 30, the action of the oxygen scavenger 50 is reduced due to contact with the liquid, and side reactions other than oxygen absorption occur. Can be suppressed. In addition to the oxygen scavenger 50, electronic parts and devices such as sensors that dislike water wetting can be installed as required, and measurement data from the sensors can be obtained and the device driven via wireless communication. Can be executed. Such sensors and devices are introduced into the culture tank 13 after being sterilized as necessary.

  Next, the microorganism culture system 300 of the present invention will be described with reference to FIG. In addition, the following embodiment is only an example which actualized this invention, and it cannot be overemphasized that embodiment can be changed suitably in the range which does not change the summary of this invention. Moreover, the same code | symbol is attached | subjected to the same part as the microorganism culture apparatus 4 and the culture system 10 of above-described 2nd Embodiment, and the description is abbreviate | omitted.

  FIG. 9 is a diagram showing an overview of a microorganism culture system 300 according to an embodiment of the present invention. As shown in FIG. 9, the microorganism culture system 300 includes a plurality of microorganism culture apparatuses 4 according to the second embodiment, and is configured to be able to perform culture in parallel with the plurality of microorganism culture apparatuses 4. For this reason, a branch pipe 301 is provided in the delivery pipe 16. The branch pipe 301 is a pipe that branches the flow path, and the number of branches of each culture tank 13 is formed. A supply tube 16a connected to each supply unit 31 of each microorganism culture device 4 is attached to each branch. Thereby, the liquid stored in the storage tank 11 is supplied to each of the culture tanks 13 of each microorganism culture apparatus 4. The porous body 1 installed in each microorganism culturing apparatus 4 may be smeared with the same concentration and the same type of seed source, or with different concentrations of the seed source or with different types of environmental samples. May be.

  In addition, a mixing tube for mixing antibiotics, a specific substrate, and further sterilized water supplied only to one microorganism culturing device 4 is connected to the microorganism culturing device 4 downstream of the branch tube 301. May be. By setting it as this structure, in each culture tank 13, it can culture | cultivate in parallel, using the culture solution from which a density | concentration and a composition differ.

  Hereinafter, the present invention will be described in detail based on examples. The present invention is not limited to these examples as long as the gist thereof is not exceeded.

Example 1
After adding 10% Silica gel 60 (0.063-0.2 mm) (Merck) to the 7% potassium hydroxide solution and stirring gently, warm the prepared solution to about 60 ° C. with a microwave oven. It was. Thereafter, suction filtration sterilization was performed using a 0.22 μm pore cellulose acetate + nitrocellulose membrane filter (Merck Millipore). The sterilized solution was mixed with distilled water sterilized by an autoclave at a ratio of 1: 1 (v / v) in a clean bench. Thereafter, 2% of an ortho-phosphoric acid (manufactured by Merck) solution sterilized by an autoclave was added to the mixed solution. The finally obtained mixed solution was poured into a sterilized petri dish having a diameter of 90 mm to a height of 5 mm and solidified for 20 minutes. The solidified petri dish was placed upside down and left at room temperature for 12 hours while being irradiated with ultraviolet rays in a clean bench. Thereafter, the water exuded from the silica gel was taken out with a sterilized glass top pipette. As a result, a silica gel plate was obtained as a porous material.

  Next, in the clean bench, the side and bottom surfaces of the petri dish were cut out to form portions where the gel was exposed. Thereafter, using a culture apparatus configured in the same manner as the microorganism culture apparatus 4 described in the above embodiment, a mounting table in which appropriate legs are attached with screws to adjust the height in a sterilized culture tank The petri dish was placed on the sterilized tube, and the sterilized tubes were connected to the supply part and the discharge part of the culture tank. The other end of each of the tubes is connected to a storage bottle for storing the culture solution and a drainage bottle for storing the drainage, and a supply pump (trade name MINIPULS evolution, manufactured by Gilson) so that the liquid can be fed. Both tubes were attached to a discharge pump (trade name MINIPULS evolution, manufactured by Gilson).

  In the storage bottle, 0.001% resazurin (7-Hydroxy-3H-phenoxazin-3-one 10-oxide) Sodium Salt solution (manufactured by Wako Pure Chemical Industries) is mixed with distilled water and colored blue. Water was introduced, and both the supply pump and the discharge pump were driven to supply colored water into the culture tank. As a result, it was observed that the produced silica gel plate was colored, and it was confirmed that water was supplied (liquid supply and discharge) to the silica gel plate. Moreover, the liquid level could be formed at a position slightly lower than the surface of the gel.

(Example 2)
Next, the petri dish with a hole which has a silica gel plate similarly to what was produced in Example 1 was produced, and this petri dish was mounted on the mounting base in a culture tank using the culture apparatus similar to the said Example 1. FIG. The stored matter in the storage bottle was distilled water, and the pump was driven to flow distilled water, and water was passed while forming a liquid surface at a position slightly lower than the surface of the gel. Under this condition, a blue colored solution is applied to the surface of the silica gel plate with 0.001% resazurin (7-Hydroxy-3H-phenoxazin-3-one 10-oxide) Sodium Salt (Wako Pure Chemical Industries, Ltd.). As a result of observation, it was observed that the color of the plate surface was colored after 30 minutes and the wastewater was colored blue. Thus, it was shown that the substance can be transferred from the surface of the silica gel plate by the liquid that flows through (substance exchange is possible).

(Example 3)
Using an environmental sample (activated sludge in the aeration tank of Toyohashi University of Technology) as a microbial source, 0.5 ml of the sample was collected and 10 ^-1 with 4.5 ml of 10 mM sodium dihydrogen phosphate (Wako Pure Chemical Industries, Ltd.) solution. Dilute serially to ^10-5 times. Among them, 10 ⁻⁵ times suspension was used as a seeding source. As in Example 1, a petri dish containing a silica gel plate was prepared, and 50 μL of the prepared suspension was smeared on the silica gel plate, and then set in the culture apparatus as in Example 1. Moreover, the mounting base which mounts a petri dish adjusted the height by attaching the leg part of suitable height (22 mm) with a screw stop.

  Next, a storage bottle storing a liquid medium adjusted to pH 7.0 by mixing 0.01% tryptone, 0.005% yeast extract and 0.005% sodium chloride was prepared. Thereafter, the storage bottle containing the liquid medium was sterilized by autoclaving. Then, the supply pump is driven to supply the liquid medium at a flow rate of 1.25 ml / min, and the medium is discharged at a flow rate of 1.30 ml / min with the discharge pump, so that the medium is lowered 2-3 mm below the surface of the silica gel plate. In a state where a liquid surface was formed, continuous culture was performed on a silica gel plate at 28 ° C. for 3 days. As a result, it was confirmed that colonies were generated only on the surface of the gel plate after 2 days of culture and were not lost even after 3 days of culture. Thereby, even if it culture | cultivated supplying a culture solution to a silica gel (namely, porous body), it was shown that it can isolate | separate and culture in the state fixed to the fixed position of the porous body surface, without microbial colony disappearing. The 330 colonies formed on the plate were small and round with a diameter of about 0.5 mm.

1 Porous body 2, X Petri dish (container)
3, 4, 100 Microbe culture apparatus 3a, 30 Culture chamber 3b, 31 Supply section (part of the flow path)
3c, 32 discharge part (part of flow path)
11 Storage tank 12 Supply pump 13 Culture tank (outer tank)
13a Storage tank (housing)
13b Lid 13d Sealing rubber (sealing material)
14 Discharge pump 16 Delivery pipe (part of flow path)
17 Petri dish cover (part of locking tool, part of holding part)
18 Fixing screws (part of locking tool, part of holding part)
20,200 mounting table (part of holding part)
21 Superstructure (pedestal)
21c, 210c Through hole 22 Leg (support, part of adjusting means)
23 Countersunk screw (fixing tool, part of adjusting means)
220 Nut (fixing tool, part of adjusting means)
230 Volts (support, part of adjusting means)
40 Fixture (Jig)
50 Oxygen absorber 300 Microbial culture system 301 Branch pipe (part of flow path)

Claims (13)

An apparatus for culturing microorganisms,
A culture chamber configured to have a pore that is configured to allow liquid to pass therethrough and communicate with the surface, and in which the pore on the upper surface portion is formed with a pore diameter smaller than that of the microorganism;
A flow path for supplying and discharging liquid to the culture chamber;
A holding unit for holding the porous body in the culture chamber;
The liquid level formed by supplying the liquid from the flow path to the culture chamber is the same height as the upper surface of the porous body held by the holding unit or near the upper surface of the porous body and below the upper surface of the porous body. And adjusting means for adjusting the positional relationship between the porous body and the liquid surface,
Supplying liquid to the upper surface of the porous body through the pores of the porous body in a state where the liquid level is formed at the same height or slightly lower than the upper surface of the porous body while supplying liquid continuously or intermittently to the culture chamber And culturing microorganisms on the upper surface of the porous body. The holding portion has a pedestal on which the porous body is placed, and a locking tool that detachably locks the porous body on the pedestal,
The adjusting means includes a support provided in the holding portion, to which the pedestal is detachably attached, and a fixture for fixing the support to the pedestal. 2. The microorganism culture apparatus according to claim 1, wherein the positional relationship between the upper surface of the porous body and the liquid surface is adjusted by changing the height. The holding portion has a pedestal on which the porous body is placed, and a locking tool that detachably locks the porous body on the pedestal,
The adjustment means includes a support provided on the holding portion and to which the pedestal is attached, and a fixture that fixes the pedestal so that the pedestal can move in the vertical direction. The pedestal moves in the vertical direction. The microorganism culture apparatus according to claim 1, wherein the positional relationship between the upper surface of the porous body and the liquid surface is adjusted.   The microorganism culture apparatus according to claim 2 or 3, wherein the pedestal includes a through hole through which the liquid supplied to the culture chamber passes in the thickness direction of the pedestal. A container containing the porous body, the container having a through-hole communicating with the inside and outside of the container;
The microorganism holding apparatus according to any one of claims 1 to 4, wherein the holding unit holds the container in the culture chamber.   The microorganism culture apparatus according to any one of claims 1 to 5, wherein the porous body has an upper surface formed of gel.   7. The microorganism culturing apparatus according to claim 6, wherein the porous body is entirely formed of a gel. It has a bottomed casing that opens upward, and a lid that closes the opening, and includes an outer tank that encloses the culture chamber with a gap between the casing inner wall,
The microorganism culture apparatus according to any one of claims 1 to 7, wherein the flow path passes through a wall of the outer tank and is directly connected to the culture chamber.   The outer tub includes a jig that fixes the lid in a closed state, and a sealing material that is provided at an opening portion of the housing and prevents entry of outside air when the lid is fixed by the jig. The microorganism culture apparatus according to claim 8, wherein the microorganism culture apparatus is configured to be sealable. An oxygen scavenger is provided in the internal space of the outer tub,
The microorganism culture apparatus according to claim 9, wherein the outer tank is made an anaerobic atmosphere by the oxygen scavenger. A plurality of the microorganism culture devices according to any one of claims 1 to 10,
A storage tank for storing a liquid to be supplied to the plurality of microorganism culture apparatuses;
A supply pump for supplying liquid stored in the storage tank to a flow path connected to each of the plurality of microorganism culture devices;
A discharge pump for discharging the liquid from each of the microorganism culture devices,
A microorganism culture system that performs culture in parallel in each of the microorganism culture apparatuses.   A sample containing microorganisms is placed on the upper surface of a porous body that has a pore that is configured to allow liquid to pass therethrough and communicates with the surface and has a pore diameter smaller than that of the microorganism. A medium is continuously supplied to the body, and the medium is passed in the direction intersecting the height direction inside the porous body, and the medium is supplied to the upper surface of the porous body via the pores inside the porous body. And culturing microorganisms on the upper surface of the porous body.   A plurality of the porous bodies are provided, one solution selected from a plurality of solutions diluted with different concentrations or different seeding sources is smeared on the top surface of the one porous body, and each of the porous bodies smeared with the solution The culture method according to claim 12, wherein a culture medium is supplied to the plurality of cultures in parallel.

Images