Culture vessel, culture kit, and method for sedimentation testing of microorganisms

Abstract

To provide culture vessels and culture kits that eliminate the complexity of testing procedures, and to provide a method for sedimentation testing of microorganisms.SOLUTION: The culture vessel of the present invention is a culture vessel for sedimentation testing of microorganisms, and includes a container body capable of containing a culture medium. The container body has a top, a body, and a bottom. The top has a first opening configured to receive the microorganisms dropping from the environment above the culture vessel into the container body. The top or body has a second opening configured to pour the culture medium into the container. The culture vessel further includes a first lid that closes the first opening and a second lid that closes the second opening.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a culture vessel and a culture kit used in a falling-cell test for microorganisms, and a method for performing a falling-cell test for microorganisms using the culture vessel. [Background technology]

[0002] The falling bacteria method has been known for some time as a method for testing microorganisms in the environment. The falling bacteria method, also known as the Koch method, involves leaving an agar medium (plate medium) of a certain size open for a certain period of time, culturing the microorganisms that have fallen onto the surface of the agar medium, and then testing the microorganisms using the colonies that have grown. In this method, the number of colonies that have grown is counted as the number of bacteria per certain period of time, and the colonies are picked up to check the properties of the microorganisms after cultivation. Figure 9 is a flowchart that outlines the flow of a falling bacteria test using the conventional falling bacteria method, and Figures 10(a) to (d) are schematic process diagrams showing a falling bacteria test using the conventional falling bacteria method.

[0003] In a conventional falling bacteria test using the falling bacteria method, as shown in the flowchart of FIG. 9, first, as shown in FIG. 10(a), a petri dish containing an agar medium is placed at the test location with its lid open (placement of the petri dish at the test location). Next, the petri dish is maintained at the test location with its lid open until a certain amount of time has passed since its placement at the test location. This allows microorganisms in the environment above the petri dish to fall onto the agar medium in the petri dish. Then, as shown in FIGS. 10(a) and 10(b), the petri dish lid is closed by placing the petri dish with the agar medium facing downwards against the lid, with the agar medium facing up. This allows microorganisms to be collected (collection of microorganisms). Next, the petri dish with the lid closed is placed in an incubator (not shown) set to the temperature range required for culturing the target microorganisms and maintained there for a predetermined period of time, allowing solid-state culture of the microorganisms on the agar medium in the petri dish (cultivation of microorganisms), as shown in FIG. 10(c). Next, the number of microbial colonies after cultivation is counted (colony counting). Next, as shown in Figure 10(d), colonies are picked up using a platinum loop (colony picking). Next, the picked colonies are used to perform tests such as confirming the properties of the microbial cells after cultivation (microbial testing).

[0004] As a conventional technique for culturing and testing microorganisms such as the above-mentioned falling bacteria test, for example, a technique for a film-like culture medium for culturing microorganisms is known (Patent Document 1). This technique can provide a culture medium that eliminates the labor required for culture medium preparation, has good storage properties, saves space, and reduces waste. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2007-20434 Summary of the Invention [Problem to be solved by the invention]

[0006] In conventional falling bacteria testing, as described above, it is necessary to pick up colonies in order to check the properties of the microorganisms after cultivation. The work of picking up colonies is complicated and time-consuming.

[0007] The present invention has been made in consideration of these points, and its object is to provide a culture vessel and culture kit to be used in the testing of falling microorganisms, and a method for performing the testing of falling microorganisms, which can eliminate the complexity of the testing work. [Means for solving the problem]

[0008] In order to solve the above problem, the culture vessel of the present invention is a culture vessel used for testing falling microorganisms, and is equipped with a container body capable of containing a culture medium therein, the container body having a top, a body, and a bottom, the top being provided with a first opening configured to receive the microorganisms falling from the environment above the culture vessel into the container body, the top or the body being provided with a second opening configured to pour the culture medium, and the culture vessel further being equipped with a first lid that closes the first opening and a second lid that closes the second opening.

[0009] In addition, the culture kit of the present invention is a culture kit used for testing falling microorganisms, and is characterized by comprising the above-mentioned culture vessel, a microorganism collection filter, and an adhesive material used to attach the microorganism collection filter to the area on the bottom surface inside the container body of the culture vessel where the microorganisms fall through the first opening.

[0010] In addition, the method for performing a falling bacteria test for microorganisms of the present invention is a method for performing a falling bacteria test for microorganisms using the above-mentioned culture vessel, and is characterized by comprising the steps of: placing the culture vessel at an inspection location with the first opening open; collecting microorganisms by allowing microorganisms in the environment above the culture vessel to fall into the inside of the container body through the first opening until a certain time has elapsed since the culture vessel was placed at the inspection location; supplying a liquid culture medium into the inside of the container body through the first opening or the second opening after collecting the microorganisms; obtaining a culture solution by culturing the microorganisms in the liquid culture medium supplied into the inside of the container body; removing the culture solution from the inside of the container body through the second opening to the outside of the culture vessel; and testing the microorganisms using the culture solution removed to the outside. [Effects of the Invention]

[0011] According to the present invention, the complexity of the inspection work can be eliminated. [Brief explanation of the drawings]

[0012] [Figure 1] 1(a) and 1(b) are schematic perspective views illustrating a state in which microorganisms are collected from the culture vessel according to the first embodiment and a state in which the culture medium is taken out to the outside, respectively. [Figure 2] 1 is a flowchart showing an outline of the flow of a method for performing a falling bacteria test for microorganisms according to the first embodiment. [Figure 3] 10(a) and 10(b) are schematic perspective views illustrating a state in which microorganisms are collected from a culture vessel according to a second embodiment and a state in which the culture medium is taken out to the outside, respectively. [Figure 4] 10(a) and 10(b) are schematic perspective views illustrating the initial state and the state when the culture medium is removed to the outside, respectively, of the culture kit according to the third embodiment. [Figure 5] FIG. 10 is a schematic perspective view illustrating an example of a culture vessel according to a modified example of the first embodiment. [Figure 6]FIG. 10 is a schematic perspective view illustrating an example of a culture vessel according to a modified example of the second embodiment. [Figure 7] FIG. 11 is a schematic perspective view illustrating a culture kit according to a modified example of the third embodiment. [Figure 8] 10(a) and 10(b) are schematic perspective views illustrating a second cover of a culture vessel according to a modified example of the first embodiment. [Figure 9] 1 is a flowchart showing an outline of a flow of a falling bacteria inspection using a conventional falling bacteria method. [Figure 10] 1(a) to 1(d) are schematic process diagrams showing a falling bacteria inspection using a conventional falling bacteria method. DETAILED DESCRIPTION OF THE INVENTION

[0013] Hereinafter, embodiments of the culture vessel, culture kit, and method for performing a falling bacteria test for microorganisms of the present invention will be described.

[0014] [First embodiment] First, a description will be given of a culture vessel and a method for performing a falling bacteria test for microorganisms according to the first embodiment. Figures 1(a) and 1(b) are schematic perspective views illustrating the state when collecting microorganisms in the culture vessel according to the first embodiment and the state when the culture solution is taken out, respectively.

[0015] As shown in FIGS. 1(a) and 1(b), the culture vessel 1 according to the first embodiment is a culture vessel 1 used in a falling microorganism test and includes a vessel body 2 capable of containing a culture medium therein. The vessel body 2 includes a top (upper) portion 4, a body 6, and a bottom 8. The vessel body 2 has a general shape of a flat rectangular prism, with the surface 4s of the top portion 4 and the surface 8s of the bottom 8 serving as the bottom surface and the surface 6s of the body 6 serving as the side surface. The top portion 4 and the bottom 8 have the same rectangular plate-like shape with one short side corner removed, and the body 6 has a general shape formed by joining multiple rectangular plate-like members 6p circumferentially, connecting multiple sides of the top portion 4 with corresponding sides of the bottom 8. The top portion 4 is provided with a first opening 10 configured to receive microorganisms falling from the environment above the culture vessel 1 into the vessel body 2. The first opening 10 has a rectangular shape with its short and long sides parallel to those of the top 4. Of the multiple plate-like members 6p of the body 6, the plate-like member 6p connecting the short sides of the top 4 and bottom 8 where the corners have been cut off has a neck 6n that protrudes outward from its surface 6ps. The neck 6n of the body 6 has an openable second opening 12 configured to allow the culture medium to be poured into it.

[0016] The culture vessel 1 further includes a sliding lid 20, which serves as a first lid for opening and closing the first opening 10, and slides and moves in a direction along the short sides of the first opening 10 to open and close the first opening 10. The sliding lid 20 includes a lid body 20b and a gripping portion 20g. The lid body 20b includes a rectangular, plate-shaped lid portion 20p and two sliding portions 20s extending from the two short sides of the lid portion 20p to the opposite side of the lid portion 20p. The short and long sides of the lid portion 20p are parallel to the short and long sides of the first opening 10, respectively. To enable the lid portion 20p to completely close the first opening 10, the dimensions of the short sides of the lid portion 20p are slightly longer than the short sides of the first opening 10, and the dimensions of the long sides of the lid portion 20p are the same as the long sides of the first opening 10. The sliding portion 20s has a rectangular plate shape with two short sides extending from the ends of the two long sides of the lid portion 20p in directions parallel to the long sides, and the long side of the sliding portion 20s facing the lid portion 20p is common to the short sides of the lid portion 20p. The gripping portion 20g protrudes upward from the surface of the lid portion 20p on the long side facing the second opening 12 of the lid portion 20p.

[0017] Two guide portions 2g are provided on the back surface 4r of the top portion 4 of the container body 2 to guide the two sliding portions 20s of the sliding lid 20, respectively, so that the sliding lid 20 can slide toward the second opening 12 and the opposite side in a direction along the short side of the first opening 10. Each of the two guide portions 2g extends from the end of one of the two short sides of the first opening 10 of the top portion 4 on the second opening 12 side in a direction parallel to the short side to the short side of the top portion 4 opposite the second opening 12. The guide portions 2g are shaped like a rectangular plate whose short sides are parallel to the long sides of the first opening 10 and whose long sides are parallel to the short sides of the first opening 10. The guide portions 2g are positioned so that the entire short side of the first opening 10 overlaps with the long side of the guide portion 2g on the first opening 10 side when the top portion 4 is viewed in plan. The long side of the guide portion 2g opposite to the first opening 10 is fixed to the back surface 4r of the top portion 4. The sliding lid 20 has the lid body 20b disposed inside the container body 2 so that the two sliding portions 20s are sandwiched between the back surface 4r of the top portion 4 and the two guide portions 2g, respectively, and the gripping portion 20g protrudes to the outside of the container body 2 through the first opening 10.

[0018] The culture vessel 1 further includes a cap 16 (second lid) that can be attached to and detached from the second opening 12 so as to open and close the second opening 12. A screw thread (not shown) is provided on the inner portion of the cap 16, and a screw thread 6nc is also provided on the outer portion of the neck portion 6n where the second opening 12 is provided, and these screw threads interlock with each other. After the cap 16 is placed on the neck portion 6n, the screw threads of the cap 16 are engaged with the screw threads 6nc of the neck portion 6n, and the cap 16 is wound and tightened, thereby attaching the cap 16 to the second opening 12 and closing the second opening 12. From this state, the cap 16 can be removed from the second opening 12 and opening the second opening 12 by winding and loosening the cap 16 in the reverse direction.

[0019] The culture vessel 1 further includes a microorganism collection filter 30 attached to an area on the bottom surface inside the vessel body 2 where the microorganisms fall through the first opening 10. The microorganism collection filter 30 is a membrane filter with a filter pore size of 0.45 μm or less, and is made of a material that does not have an adverse effect on microorganisms, and has a structure that does not have an adverse effect on microorganisms.

[0020] In the method for performing a falling bacteria test for microorganisms according to the first embodiment, the culture vessel 1 according to the first embodiment is used to perform a falling bacteria test for microorganisms. Fig. 2 is a flowchart that schematically shows the flow of the method for performing a falling bacteria test for microorganisms according to the first embodiment.

[0021] In the method according to the first embodiment, as shown in the flowchart of FIG. 2, first, as shown in FIG. 1(a), a cap 16 is attached to the second opening 12 of the culture vessel 1 to close the second opening 12, and then the sliding lid 20 is slid and moved to the opposite side of the second opening 12 to open the first opening 10, and the culture vessel 1 is placed at the test location (placement of the culture vessel at the test location). Next, the culture vessel 1 is maintained at the test location until a certain amount of time has elapsed since the culture vessel 1 was placed at the test location. As a result, microorganisms in the environment above the culture vessel 1 are dropped through the first opening 10 onto the microorganism collection filter 30 inside the container body 2, and are collected by the microorganism collection filter 30. Then, the sliding lid 20 is slid and moved toward the second opening 12 to temporarily close the first opening 10. As a result, microorganisms are collected inside the container body 2 (collection of microorganisms).

[0022] Next, the sliding lid 20 is slid to the opposite side from the second opening 12 to open the first opening 10 again, and a liquid medium (not shown) is supplied into the container body 2 through the first opening 10 (supply of liquid medium). At this time, for example, about ¼ of the total volume of the interior of the container body 2 is supplied into the container body 2 so that the entire microorganism collection filter 30 is immersed in the liquid medium. By supplying the liquid medium, the microorganisms captured by the microorganism collection filter 30 are separated from the microorganism collection filter 30 and become suspended in the liquid medium.

[0023] Next, the sliding lid 20 is slid toward the second opening 12 to close the first opening 10, and the culture vessel 1 is placed in an incubator (not shown) set to the temperature range required for culturing the microorganisms to be cultured and maintained for a predetermined period of time, thereby culturing the microorganisms in the liquid medium supplied inside the vessel body 2 to obtain a culture solution (cultivation of microorganisms).

[0024] 1(b), the cap 16 is removed from the second opening 12 of the culture vessel 1 to open the second opening 12, and the culture medium is removed from inside the vessel body 2 through the second opening 12 to the outside of the culture vessel 1 (removal of the culture medium). In this case, for example, the culture medium is poured from inside the vessel body 2 through the second opening 12 into a test vessel (not shown) such as a centrifuge tube, and the culture medium is removed directly into the test vessel.

[0025] Next, the culture medium removed to the outside is used to test the microorganisms (microorganism testing). At this time, the microorganism testing includes confirming the properties of the microorganisms and conducting genetic testing. Examples of methods for genetic testing of microorganisms include extracting nucleic acids from the cultured microorganisms contained in the culture medium removed to a testing container such as a centrifuge tube, amplifying the nucleic acids using a PCR method or the like, and analyzing the genes of the amplified nucleic acids to identify the microorganisms. As described above, a falling cell test of microorganisms is performed using the culture container 1 according to the first embodiment.

[0026] In the culture vessel 1 according to the first embodiment, a first opening 10 is provided on the top 4 of the vessel body 2, and is configured to receive microorganisms that fall from the environment into the vessel body 2. Furthermore, a second opening 12 that can be opened and closed and configured to pour a culture medium is provided on the body 6 of the vessel body 2. Furthermore, the culture vessel 1 according to the first embodiment includes a slide lid 20 (first lid) that opens and closes the first opening 10, and a cap 16 (second lid) that can be attached and detached to the second opening 12 so that the second opening 12 can be opened and closed. Thus, in the method for performing a falling bacteria test for microorganisms according to the first embodiment, by using the culture vessel 1 according to the first embodiment, the first opening 10 is opened, and microorganisms in the environment are allowed to fall into the vessel body 2 through the first opening 10, thereby allowing the collection of microorganisms by the falling bacteria method. Furthermore, after the collection of the microorganisms, a liquid medium is supplied into the vessel body 2 through the first opening 10, and the microorganisms are cultured in the liquid medium supplied into the vessel body 2, thereby obtaining a culture medium. Furthermore, the culture medium can be removed from the inside of the container body 2 to the outside of the culture container 1 through the second opening 12, and the removed culture medium can be used to confirm the properties of the microorganisms, perform genetic testing, and other tests. Therefore, unlike conventional drop-off bacterial testing, there is no need to pick up microbial colonies after cultivation to perform tests such as confirming the properties of the microorganisms or performing genetic testing. Furthermore, since the culture medium can be poured from the inside of the container body 2 into a test container such as a centrifuge tube through the second opening 12 and the culture medium can be directly removed from the test container, there is no need to use a pipette or other tool to remove the culture medium from the inside of the container body 2 to the test container. This eliminates the complexity of the testing process. It also reduces the risk of contamination during the processing of microorganisms for testing. Furthermore, it enables testing to be performed by personnel other than those with high expertise. In addition, because liquid culture rather than solid-state culture is used for culturing microorganisms, the time required for culturing the microorganisms to grow can be shortened, thereby shortening the testing time.

[0027] Furthermore, the culture vessel 1 according to the first embodiment further includes a microorganism collection filter 30 attached to the area on the bottom surface of the interior of the vessel body 2 where microorganisms fall through the first opening 10. The microorganism collection filter 30 is a membrane filter with a filter pore size of 0.45 μm or less, made of a material that does not adversely affect microorganisms, and has a structure that does not adversely affect microorganisms. Thus, in the method for performing a falling microorganism test according to the first embodiment, when microorganisms are collected by dropping them into the interior of the vessel body 2 through the first opening 10, the microorganisms can be collected by dropping them onto the microorganism collection filter 30, preventing the collected microorganisms from being damaged. This ensures reliable cultivation of the microorganisms. Furthermore, because the microorganism collection filter 30 is attached to the area where microorganisms fall, the position of the microorganism collection filter 30 does not shift, and the microorganisms can be reliably dropped onto the microorganism collection filter 30 and collected.

[0028] [Second embodiment] Next, a culture vessel and a method for performing a falling bacteria test for microorganisms according to the second embodiment will be described, focusing on the differences from the first embodiment. Figures 3(a) and 3(b) are schematic perspective views illustrating the state when collecting microorganisms in the culture vessel according to the second embodiment and the state when the culture solution is taken out, respectively.

[0029] 3(a) and 3(b), the culture vessel 1 according to the second embodiment has a vessel body 2 similar to that of the culture vessel according to the first embodiment, except that the first opening 10 is located closer to the center of the top section 4 and that a guide section 2g is not provided. Unlike the culture vessel according to the first embodiment, the culture vessel 1 according to the second embodiment further includes a hinged lid 40, which serves as a first lid for opening and closing the first opening 10. The hinged lid 40 opens and closes the first opening 10 by rotating a lid body 40b toward and away from the top section 4 around a hinge 40h like a swing door. The hinged lid 40 has a rectangular, plate-like lid body 40b, a hinge 40h that connects one side of the lid body 40b to one side of the first opening 10 in the top section 4, and a gripping section 40g. The lid body 40b has a rectangular plate-shaped lid portion 40p and a rib 40r that is provided on the surface of the lid portion 40p facing the first opening 10 and engages with the periphery of the first opening 10 in the top portion 4. The culture vessel 1 according to the second embodiment further includes a cap 16 (second lid) similar to the culture vessel according to the first embodiment. The culture vessel 1 according to the second embodiment, like the culture vessel according to the first embodiment, further includes a microorganism collection filter 30 that is affixed to an area on the bottom surface inside the vessel body portion 2 where microorganisms fall through the first opening 10.

[0030] In the method for performing a falling bacteria test for microorganisms according to the second embodiment, the culture vessel 1 according to the second embodiment is used to perform a falling bacteria test for microorganisms. In this case, when opening the first opening 10, the lid body 40b of the hinged lid 40 is rotated toward the opposite side of the top 4 around the hinge 40h. When closing the first opening 10, the lid body 40b of the hinged lid 40 is rotated toward the top 4 around the hinge 40h, and the rib 40r of the lid body 40b engages with the periphery of the first opening 10 in the top 4. Except for these points, the falling bacteria test for microorganisms is performed in the same manner as the method for performing a falling bacteria test for microorganisms according to the first embodiment. Therefore, the culture vessel and the method for performing a falling bacteria test for microorganisms according to the second embodiment provide the same effects as those of the first embodiment.

[0031] [Third embodiment] Next, a culture kit and a method for performing a falling bacteria test for microorganisms according to the third embodiment will be described, focusing on differences from the first embodiment. Figures 4(a) and 4(b) are schematic perspective views illustrating the state when collecting microorganisms and the state when removing the culture solution from the culture kit according to the third embodiment, respectively.

[0032] As shown in FIGS. 4(a) and 4(b), a culture kit 100 according to the third embodiment includes a culture vessel 1, a microorganism collection filter 30, and an adhesive material 32 used to attach the microorganism collection filter 30 to an area on the bottom surface of the interior of the container body 2 of the culture vessel 1 where microorganisms fall through the first opening 10. The culture vessel 1 includes a container body 2 similar to that of the culture vessel according to the first embodiment, except that the first opening 10 is located closer to the center of the top 4 and that a guide portion 2g is not provided. Unlike the culture vessel according to the first embodiment, the culture vessel 1 further includes a sheet lid 50 as a first lid for opening and closing the first opening 10. The sheet lid 50 includes a rectangular lid sheet 50b large enough to cover the first opening 10 and an adhesive sticker 50s attached to the outer periphery of the attachment surface of the lid sheet 50b. The culture vessel 1 further includes a cap 16 (second lid) similar to that of the culture vessel according to embodiment 1. The microorganism collection filter 30 is similar to that of the culture vessel according to embodiment 1. The adhesive material 32 is a double-sided adhesive sheet having a base sheet 32s and two adhesive layers 32a provided on each side of the base sheet.

[0033] In the method for performing a falling microorganism test according to the third embodiment, a culture kit 100 according to the third embodiment is used to perform a falling microorganism test. In this case, before placing the culture vessel 1 at the test location, a microorganism collection filter 30 is attached to the area of ​​the bottom surface of the interior of the vessel body 2 of the culture vessel 1 where microorganisms will fall through the first opening 10 using an adhesive material (double-sided adhesive sheet) 32. When closing the first opening 10, the adhesive seal 50s of the sheet lid 50 is attached to the periphery of the first opening 10 on the surface 4s of the top 4, thereby attaching the sheet lid 50 to the surface 4s of the top 4 so as to cover the first opening 10. When reopening the first opening 10 after temporarily closing it, the adhesive seal 50s of the sheet lid 50 is peeled off from the surface 4s of the top 4, thereby peeling the sheet lid 50 from the surface 4s of the top 4. Except for these points, the falling microorganism test is performed in the same manner as the method for performing a falling microorganism test according to the first embodiment. Therefore, the culture vessel and the method for performing a falling bacteria test for microorganisms according to the third embodiment can provide the same effects as those of the first embodiment.

[0034] [Variations] Next, modifications of the first to third embodiments will be described, focusing on differences from the first to third embodiments. Fig. 5 is a schematic perspective view illustrating a culture vessel according to a modification of the first embodiment. Fig. 6 is a schematic perspective view illustrating a culture vessel according to a modification of the second embodiment. Fig. 7 is a schematic perspective view illustrating a culture kit according to a modification of the third embodiment.

[0035] As shown in FIG. 5, the culture vessel 1 according to the modified example of the first embodiment differs from the culture vessel according to the first embodiment in that a vent hole 6h is provided in the plate-shaped member 6p on the side of the barrel 6 of the vessel body 2 opposite the cap 16 (second opening 12), and a hydrophobic filter 60 (breathable membrane material) is attached to the surface 6ps of the plate-shaped member 6p so as to cover the vent hole 6h. Furthermore, unlike the culture vessel according to the first embodiment, the culture vessel 1 further includes a non-breathable cover 70 that opens and closes the vent hole 6h. The non-breathable cover 70 is a single-sided adhesive sheet having a base sheet 70s and an adhesive layer 70a provided on one side of the base sheet 70s. The adhesive layer 70a of the non-breathable cover 70 is attached to the surface 6ps of the plate-shaped member 6p, and the non-breathable cover 70 is attached to the surface 6ps of the plate-shaped member 6p so as to cover the vent hole 6h and the hydrophobic filter 60, thereby closing the vent hole 6h. The ventilation hole 6h can be opened by peeling the adhesive layer 70a of the non-air-permeable cover 70 from the surface 6ps of the plate-shaped member 6p and peeling the non-air-permeable cover 70 from the surface 6ps of the plate-shaped member 6p. Except for these points, the culture vessel 1 is the same as the culture vessel according to the first embodiment.

[0036] A method for performing a microorganism dropping test using the culture vessel 1 according to a modification of the first embodiment achieves the same effects as the first embodiment. Furthermore, in this method, the culture vessel 1 is placed at the test location, and when microorganisms in the environment above the culture vessel 1 are dropped into the container body 2 through the first opening 10, the vent 6h is opened, allowing air to flow from the inside of the container body 2 to the outside through the vent 6h. This makes it easier for the microorganisms to drop into the container body 2 through the first opening 10. This ensures that the microorganisms in the environment are collected. Furthermore, when collecting aerobic bacteria as microorganisms, opening the vent 6h when culturing the microorganisms in a liquid medium supplied to the container body 2 allows air to be introduced into the liquid medium from the outside of the container body 2 through the vent 6h without allowing the liquid medium to flow out from the inside of the container body 2 through the vent 6h. This promotes the cultivation of aerobic bacteria. On the other hand, when collecting anaerobic bacteria as microorganisms and culturing the microorganisms in a liquid culture medium supplied inside the container body 2, by attaching a non-breathable cover 70 to the surface 6ps of the plate-shaped member 6p to close the ventilation hole 6h, it is possible to prevent air from being drawn into the liquid culture medium from outside the container body 2 through the ventilation hole 6h, thereby avoiding the inhibition of the cultivation of the anaerobic bacteria.

[0037] As shown in FIG. 6 , unlike the culture vessel according to the second embodiment, the culture vessel 1 according to the second embodiment does not include a microorganism collection filter 30. Except for this, the culture vessel 1 is similar to the culture vessel according to the second embodiment. In a method for performing a falling microorganism test using the culture vessel 1 according to the second embodiment, unlike the second embodiment, when collecting microorganisms by dropping them into the interior of the container body 2 through the first opening 10, the microorganisms are not collected by dropping them onto the microorganism collection filter 30, but are instead collected by dropping them onto the bottom surface of the interior of the container body 2. Even in this case, similar to the method for performing a falling microorganism test according to the second embodiment, after collecting the microorganisms, a liquid medium is supplied into the interior of the container body 2 through the first opening 10, and the microorganisms can be cultured in the liquid medium. This eliminates the complexity of the testing process. It also reduces the risk of contamination. Furthermore, it enables testing to be performed by operators other than those with high expertise. Furthermore, because liquid culture is performed instead of solid-state culture, the testing time can be shortened.

[0038] As shown in FIG. 7 , a culture kit 100 according to a modified example of the third embodiment includes a culture vessel 1 different from the culture kit according to the third embodiment. The culture vessel 1 has a first opening 10 located closer to the cap 16 (second opening 12) of the top section 4. A vent 6h is located on the top section 4 of the container body 2, opposite the cap 16 (second opening 12), and a hydrophobic filter 60 (breathable membrane material) is attached to the surface 4s of the top section 4 so as to cover the vent 6h. The culture vessel 1 further includes a non-breathable cover 70 that opens and closes the vent 6h. The non-breathable cover 70 is a single-sided adhesive sheet having a base sheet 70s and an adhesive layer 70a provided on one side of the base sheet 70s. The adhesive layer 70a of the non-breathable cover 70 is attached to the surface 4s of the top section 4, and the non-breathable cover 70 is attached to the surface 4s of the top section 4 so as to cover the vent 6h and the hydrophobic filter 60, thereby closing the vent 6h. The ventilation hole 6h can be opened by peeling the adhesive layer 70a of the non-breathable cover 70 from the surface 4s of the top 4 and peeling the non-breathable cover 70 from the surface 4s of the top 4. Except for these points, the culture vessel 1 is similar to the culture vessel according to the third embodiment. In the method for performing a dropping bacteria test for microorganisms using the culture kit 100 according to the modified example of the third embodiment, the same effects as those of the third embodiment can be obtained, as well as the same effects as those of the modified example of the first embodiment.

[0039] Next, the configurations of the culture vessel and culture kit according to the embodiment, and the method for performing a falling bacteria test for microorganisms will be described in further detail.

[0040] 1.Culture container A culture vessel according to an embodiment is a culture vessel used in a dropping bacteria test for microorganisms, and includes a container body capable of holding a culture solution therein, the container body having a top, a body, and a bottom, the top being provided with a first opening configured to receive the microorganisms dropping from the environment above the culture vessel into the container body, the top or the body being provided with a second opening configured to pour the culture solution into, and the culture vessel further including a first lid for closing the first opening and a second lid for closing the second opening. That is, the culture vessel is a culture vessel used for both liquid culture of microorganisms and a dropping bacteria test.

[0041] The material of the container body is not particularly limited, but examples thereof include PET (polyethylene terephthalate), etc. The shape of the container body is not particularly limited, but it is preferable that it is not bulky and easy to carry and place at the testing site, and specifically, for example, it is preferable that the bottom surface is flat and the height of the body (the distance between the top surface and the bottom surface) is low.

[0042] The first lid is not particularly limited as long as it closes the first opening. For example, it may be a lid that opens and closes the first opening. Specific examples include a sliding lid that opens and closes the first opening by sliding, such as the sliding lid of the first embodiment; a hinged lid that opens and closes the first opening by rotating the lid body around a hinge, such as the hinged lid of the second embodiment; and a sheet lid that is attached to the surface of the top portion so as to cover the first opening and is peeled off, such as the sheet lid of the third embodiment. The first lid may or may not have a leak-proof structure that prevents the culture medium from leaking from the inside of the container body through the first opening to the outside, but a leak-proof structure is preferred. An example of a leak-proof structure is a lid that has ribs on the lid body, such as the hinged lid of the second embodiment, and the ribs fit around the periphery of the first opening in the top portion to prevent leakage of the culture medium. Other examples include a sheet lid according to the third embodiment, which has a rectangular lid sheet large enough to cover the first opening and an adhesive seal attached to the outer periphery of the lid sheet's adhesive surface, with the adhesive seal adhering to the periphery of the first opening on the surface of the top part to prevent leakage of the culture medium. Examples of sliding lids include, but are not limited to, those made of PET or the like. Examples of opening lids include, but are not limited to, those in which the lid portion, hinge, and grip portion of the lid are made of PET or the like, and the lid ribs are made of rubber or the like. Examples of sheet lids include, but are not limited to, those in which the lid sheet is made of PET or the like, and the adhesive seal is made of an adhesive containing at least one type selected from the group consisting of silicone-based resins and fluorine-based resins.

[0043] The second lid is not particularly limited as long as it closes the second opening, but for example, a lid that can be removed from the second opening to open it is preferred, and among these, a cap that can be attached to and removed from the second opening, like the second lids of the first and second embodiments, is preferred. This is because the second opening can be closed by attaching the cap, and the second opening can be opened by removing the cap. The material of the cap is not particularly limited, but examples include PET (polyethylene terephthalate).

[0044] 8(a) , the second lid may be, for example, a sealing lid 17 that is attached to the surface 4s of the top portion 4 or the surface 6s of the body portion 6 (e.g., the surface 6ns of the neck portion 6n) to close the second opening 12, and that can be removed from the second opening 12 by peeling it off the surface 4s of the top portion 4 or the surface 6s of the body portion 6. The sealing lid 17 has a base sheet 17s large enough to cover the second opening 12 and an adhesive 17a provided on the outer periphery of one side of the base sheet 17s, and is a lid that is adhered to the periphery of the second opening 12 on the surface 4s of the top portion 4 or the surface 6s of the body portion 6 (e.g., the surface 6ns of the neck portion 6n) with the adhesive 17a to prevent leakage of the culture medium. An example of the sealing lid 17 is an aluminum sealing lid in which the base sheet 17s is made of aluminum foil. 8(b), the second lid may be a lid member 6c that is integrally formed with the top portion 4 or the body portion 6 (e.g., neck portion 6n) and that can be removed from the second opening 12 by cutting it off from the body portion 6 (e.g., neck portion 6n) along a slit 6t. The material of the lid member 6c is not particularly limited, but is usually the same as the material of the container body portion 2.

[0045] Preferably, the culture vessel further comprises a microorganism collection filter disposed on the bottom surface of the interior of the vessel body in an area where the microorganisms fall through the first opening. The microorganism collection filter is a porous membrane capable of collecting microorganisms, and is not particularly limited as long as it is made of a material that does not adversely affect microorganisms and has a structure that does not adversely affect microorganisms. For example, a membrane filter is preferred, and among these, a membrane filter with a filter pore size of 0.45 μm or less is preferred. This is because a filter with a pore size of 0.45 μm or less easily collects microorganisms. For example, a circular membrane filter with a diameter of 47 mm is preferred as the microorganism collection filter. Specific examples of membrane filters include Merck's Durapore (a PVDF (polyvinylidene fluoride) membrane filter), Merck's MF-Millipore (an MCE (cellulose mixed esters) membrane filter), Merck's hydrophobic PTFE (a PTFE (polytetrafluoroethylene) membrane filter), Merck's Mitex (a PTFE membrane filter), Merck's LCR (a PTFE membrane filter), Merck's Omnipore (a PTFE membrane filter), Merck's Millipore Express (a PES (polyethersulfone) membrane filter), and Merck's Isopore (a PC (polycarbonate) membrane filter).

[0046] Furthermore, from the viewpoint of being made of a material that does not adversely affect microorganisms and having a structure that does not adversely affect microorganisms, it is preferable that the microorganism collection filter does not contain antibacterial substances, such as a metal ion coating. Furthermore, it is preferable that the microorganism collection filter is made of a material that does not inhibit the culture of microorganisms in a liquid medium, and from this viewpoint, for example, a membrane filter made of a cellulose mixed ester is preferred. Furthermore, it is preferable that the microorganism collection filter is made of a material to which microorganisms do not adhere, as this can promote the culture of microorganisms, and from this viewpoint, for example, a PVDF membrane filter, a PET (polyethylene terephthalate) membrane filter, etc. are preferred.

[0047] A preferred culture vessel has the microorganism collection filter attached to the region. This is because the microorganisms can be reliably dropped onto the microorganism collection filter and captured. A preferred culture vessel has the microorganism collection filter attached to the region with an adhesive, the adhesive containing at least one selected from the group consisting of silicone-based resins and fluorine-based resins. Silicone-based resins and fluorine-based resins are highly stable adhesive materials, and such adhesives are unlikely to dissolve in the liquid medium and adversely affect the culture.

[0048] A preferred culture vessel is one in which, for example, a vent hole is provided in at least one of the top and body portions, and a breathable membrane material that is permeable to air but not liquid, is provided to cover the vent hole, as in the culture vessels according to the modifications of the first and third embodiments. This is because microorganisms can easily fall into the interior of the container body through the first opening, ensuring reliable collection of microorganisms from the environment. Furthermore, when aerobic bacteria are collected as microorganisms and cultured in a liquid medium, this can promote the cultivation of the aerobic bacteria. The breathable membrane material is not particularly limited as long as it is permeable to liquid but permeable to air, but is preferably, for example, a hydrophobic filter. Here, "hydrophobic filter" refers to a hydrophobic porous membrane that is permeable to liquid but not to air. The hydrophobic filter is not particularly limited as long as it is such a membrane, but examples include PTFE (polytetrafluoroethylene) membrane filters and polycarbonate track-etched (PCTE) membrane filters.

[0049] Among culture vessels equipped with the above-mentioned ventilation holes and breathable membrane material, those that further include a non-breathable cover that opens and closes the ventilation hole and has airtight properties are preferred. This is because, when anaerobic bacteria are collected as microorganisms and cultured in a liquid medium, closing the ventilation hole with the non-breathable cover can prevent the culture of the anaerobic bacteria from being inhibited. The non-breathable cover is not particularly limited as long as it can open and close the ventilation hole. For example, a single-sided adhesive sheet having a base sheet and an adhesive layer provided on one side of the base sheet, as in the non-breathable covers of the modified examples of the first and third embodiments, is preferred. The material of the base sheet of the non-breathable cover is not particularly limited, but examples include the same material as the material of the container body.

[0050] 2. Culture Kit The culture kit according to the embodiment is a culture kit used for testing for falling microorganisms, such as the culture kit according to the third embodiment, and includes a culture vessel, a microorganism collection filter, and an adhesive material used to attach the microorganism collection filter to an area on the bottom surface of the interior of the vessel body of the culture vessel where the microorganisms fall through the first opening. The culture vessel included in the culture kit is a culture vessel according to the embodiments that does not include a microorganism collection filter.

[0051] The adhesive material is not particularly limited, but preferably contains an adhesive containing at least one selected from the group consisting of silicone-based resins and fluorine-based resins as an adhesive for attaching the microorganism collection filter to the above-mentioned region. This is because silicone-based resins and fluorine-based resins are adhesive materials with high stability and are unlikely to adversely affect the culture. Among other things, the adhesive material is preferably a double-sided adhesive sheet having a base sheet and two adhesive layers provided on each side of the base sheet, with the two adhesive layers containing an adhesive.

[0052] 3. How to perform a microbial fall test A method for performing a falling bacteria test for microorganisms according to an embodiment is a method for performing a falling bacteria test for microorganisms using a culture vessel according to an embodiment, and includes the steps of: placing the culture vessel at an inspection location with the first opening open; collecting microorganisms by allowing microorganisms in the environment above the culture vessel to fall into the inside of the container body through the first opening until a certain time has passed since the culture vessel was placed at the inspection location; supplying a liquid culture medium into the inside of the container body through the first opening or the second opening after collecting the microorganisms; obtaining a culture solution by culturing the microorganisms in the liquid culture medium supplied into the inside of the container body; removing the culture solution from the inside of the container body through the second opening to the outside of the culture vessel; and testing the microorganisms using the culture solution removed to the outside.

[0053] Although the method for performing a falling microorganism test is not particularly limited as long as it is the above-mentioned method, a preferred method is to perform a falling microorganism test using a culture vessel further comprising a microorganism collection filter placed in the area where the microorganisms fall through the first opening on the bottom surface of the interior of the container body, and among these, a method using a culture vessel to which a microorganism collection filter is attached in the area is preferred, and a method using a culture vessel to which the microorganism collection filter is attached in the area is particularly preferred, and the adhesive is at least one selected from the group consisting of silicone-based resins and fluorine-based resins is particularly preferred. Furthermore, a preferred method for performing a falling microorganism test is to perform a falling microorganism test using a culture vessel having an air vent in at least one of the top and body portions and an air-permeable membrane material that is liquid-proof but air-permeable, covering the air vent.

[0054] The microorganisms that are the subject of the falling bacteria test are not particularly limited, and are generally known as microorganisms that are the subject of the falling bacteria test, but examples include fungi such as mold, bacteria, etc.

[0055] In the step of collecting microorganisms, the second opening of the container body of the culture vessel may be closed and the microorganisms in the environment may fall into the container body through the first opening, or the second opening of the container body of the culture vessel may be open and the microorganisms in the environment may fall into the container body through the first opening. In this case, air flows from the inside of the container body to the outside through the second opening, making it easier for the microorganisms to fall into the container body through the first opening.

[0056] In the process of culturing microorganisms in a liquid medium supplied to the interior of the container body to obtain a culture solution, the microorganisms may be cultured in the liquid medium with both the first and second openings closed, or at least one of the first and second openings open. In this case, air can be introduced into the liquid medium from outside the container body through at least one of the first and second openings, thereby promoting the cultivation of aerobic bacteria. In this case, the culture vessel must be placed in a way that prevents the liquid medium from leaking from the interior of the container body to the outside through the open one of the first and second openings.

[0057] The above provides a detailed description of embodiments of the culture vessel and culture kit of the present invention, as well as a method for conducting a falling-cell test for microorganisms. However, the present invention is not limited to the above-described embodiments, and various design modifications can be made without departing from the spirit of the present invention as set forth in the claims. [Explanation of symbols]

[0058] 1 Culture vessel 2. Container body 4 Heavenly 6. Torso 6p Plate-shaped member 6n Neck 6h Ventilation 6c Lid member 6t notch 8 bottom 10 First opening 12 Second opening 16 Cap 17 Sealing lid 20 Slide lid 30 Microorganism collection filter 32 Adhesive material (double-sided adhesive sheet) 40 Opening lid 50 Sheet Lid 60 Hydrophobic Filter 100 Culture Kits

Claims

1. A culture vessel used for testing falling bacteria of microorganisms, a container body capable of containing a culture solution therein; The container body has a top portion, a body portion, and a bottom portion, The top portion is provided with a first opening configured to receive the microorganisms dropping from the environment above the culture vessel into the vessel body, The top portion or the body portion is provided with a second opening configured to pour the culture medium, The culture vessel further includes a first lid that closes the first opening and a second lid that closes the second opening.

2. 2. The incubation vessel according to claim 1, wherein the second lid is a cap that can be attached to and detached from the second opening.

3. 2. The culture vessel according to claim 1, further comprising a microorganism collection filter disposed in an area on the bottom surface inside the vessel body where the microorganisms fall through the first opening.

4. The culture vessel according to claim 3, characterized in that the microorganism collection filter is attached to the region with an adhesive, and the adhesive contains at least one type selected from the group consisting of silicone-based resins and fluorine-based resins.

5. The culture vessel according to claim 1, characterized in that an air vent is provided in at least one of the top portion and the body portion, and a breathable membrane material having the property of not allowing liquid to pass through but allowing air to pass through is provided to cover the air vent.

6. The culture vessel according to claim 5, further comprising a non-air-permeable cover that opens and closes the ventilation hole and has air-blocking properties.

7. A culture kit used for testing microorganisms, A culture kit comprising: a culture vessel according to any one of claims 1, 2, 5, and 6; a microorganism collection filter; and an adhesive material used to attach the microorganism collection filter to an area on the bottom surface of the interior of the vessel body of the culture vessel where the microorganisms fall through the first opening.

8. 8. The culture kit according to claim 7, wherein the adhesive material comprises an adhesive containing at least one selected from the group consisting of silicone-based resins and fluorine-based resins.

9. A method for performing a falling cell test of microorganisms using the culture vessel according to any one of claims 1 to 6, placing the culture vessel at an inspection location with the first opening open; collecting microorganisms by dropping microorganisms in the environment above the culture vessel through the first opening into the vessel body until a certain time has elapsed since the culture vessel was placed at the inspection location; After collecting the microorganisms, supplying a liquid culture medium into the container body through the first opening or the second opening; obtaining a culture solution by culturing the microorganism in the liquid medium supplied to the inside of the container body; removing the culture solution from inside the vessel body to outside the culture vessel through the second opening; and testing the microorganism using the culture medium removed to the outside.

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