JPH0774771B2 - Sampling device for aseptic sampling of seabed samples - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sampler for aseptically collecting a sample of microorganisms or the like existing on the seabed, particularly mud and sand on the deep seabed.

[0002]

2. Description of the Related Art When collecting a sample of microorganisms existing in mud, sand, etc. on the deep sea floor with a water depth of several thousand meters,
In order to obtain a sample that remains on the seabed while diving in a submersible vessel and moving the sample container sampled at the seabed to the top of the mother ship, maintain equal pressure inside and outside the sample container and It is important to prevent destruction and to keep the sample sterile to prevent external contamination.

FIG. 3 is a schematic sectional view showing a conventional sampler. The inside of the storage container 3 having the opening / closing lid 1 and the stopper plate 2 has a structure in which the sample collector 4 is inserted and fixed. The sample collector 4 has a cylindrical body 5, a sample container 6 fitted in the cylindrical body 5, and a grip portion 7 formed on the upper part of the cylindrical body 5, and the sample container 5 has a sample container 5 at one end thereof. A sampling port 8 is formed and a check valve 9 is provided inside. A pressure equalizing hole 10 that communicates the inside and outside of the sample container 6 is formed in the grip portion 7. Further, the opening / closing lid 1 is provided with a filter portion 1a.

The storage container 3 is attached to the outside of the submersible, and when taking a sample of the seabed, a person inside the submersible operates the manipulator to remove the stopper plate 2 and open the lid 1. Then, when the manipulator grips the grip portion 7 of the sample collector 4 and inserts it into the seabed, the mud or sand on the seabed enters the sample container 6 from the sample collection port 8 of the sample container 6 through the check valve 9. invade. When the collection is completed, the sample collector 4 is inserted and set in the storage container 3 by the manipulator, then the opening / closing lid 1 is closed and fixed with the stopper plate 2, and the sample container 6 is recovered by moving to the mother ship on the ocean by a submersible. However, it will be used for subsequent tests and research.

[0005]

However, in the above-mentioned conventional sample collector, it is necessary to provide the opening / closing lid 1 of the storage container 3 with the filter portion 1a in order to maintain the sample in a sterile state. There is a problem that the whole becomes large and only one or two submarines can be mounted on the submersible, and therefore only one or two samples can be obtained in one diving. Further, in order to open and close the storage container 3, the manipulator is used to attach / detach the opening / closing lid 1 and the stopper plate 2, but this manipulator operation requires skill and it is difficult to confirm whether or not the opening / closing lid 1 is securely closed. Yes, and therefore has problems with its sealability.

The present invention solves the above-mentioned problems, and by maintaining a sterile state of a sample with a simple structure, it is possible to reduce the size and weight of the sample and to collect a large number of samples. It is an object of the present invention to provide a sample collector that can simplify the operation.

[0007]

Therefore, a sample collector for aseptically collecting a seabed sample according to the present invention comprises a cylindrical container 16 and an O-ring 32 fitted in the container 16. Cylindrical collector body 17 and collector body 17
A sample container 19 which is detachably attached to the lower part of the sample container, sample collection ports 21 and 33 formed at least on the lower end of the sample container 19 and on the side part of the sampler body 17, and on the upper part of the sampler body 17. Bacteria blocking member 27 provided, lid member 30 for sealing the upper part of the collector body 17, and lid member 3
It is characterized in that it is provided with a pressure equalizing hole 31 which is formed inside 0 and connects the inside and outside of the collector main body 17 to each other.

As an embodiment, the storage container 16 of the plurality of submarine sample collectors 15 may be fixed to the collector mounting box 13 fixed near the manipulator of the submersible 11, or the collector mounting box. 13 may be filled with water, or a heat insulating material 20 made of a foamable elastic material may be provided on the outer periphery of the storage container 16. It should be noted that the numbers added to the above-mentioned configurations are for comparison with the drawings in order to facilitate understanding of the present invention, and the configurations of the present invention are not limited thereby.

[0009]

In the present invention, for example, as shown in FIGS. 1 and 2, when collecting a sample of the seabed, the upper part of the collector body 17 is grasped and inserted into a desired position on the seabed, and the sample container 19 is inserted. Inject the sample from the sample collection ports 21 and 33,
After collecting the sample, the collector main body 17 is fitted into the storage container 16. When ascending to the sea, the filter 27 and the pressure equalizing hole 3 in the sample container 19 are changed by pressure and temperature changes.
Although seawater flows in and out through the sample container 1, bacteria in the seawater from the outside are blocked by the filter 27 and the O-ring 32, so that the inside of the sample container 19 can be maintained in a sterile state, and the sample container 19 can be kept sterile. The internal microorganisms can be prevented from flowing out.

[0010]

Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 is a perspective view of the submarine sampler of the present invention mounted on a submersible, FIG. 2 shows one embodiment of the submarine sampler of the present invention, FIG. 2 (A) is a sectional view, and FIG. 2 (B). 2A is a cross-sectional view as seen in the direction of the arrow along the line BB in FIG. 2A, FIG. 2C is a cross-sectional view similar to FIG. 2B showing a modified example of the sampling port,
FIG. 2D is a partial cross-sectional view showing a modified example of the sample container.

In FIG. 1, a submersible 11 is of a type capable of submersing to the ocean floor from a mother ship on the sea as a base, and one or two manipulators 1 are provided at the front of the submersible 11.
2, a sampler mounting box 13 is fixed near the manipulator 12, and a plurality of sample samplers 15 according to the present invention can be mounted in the sampler mounting box 13. As an example, the size of the sampler 15 is about 5c in diameter.
It has a height of about 25 cm and a weight of about 1.7 kg.

In FIG. 2 (A), a sample collector 15 according to the present invention is made of stainless steel and has a cylindrical storage container 16 with a bottom.
And a cylindrical stainless steel collector body 17 fitted in the storage container 16, and a sample container 19 which is detachably attached to the lower portion of the collector body 17 by means such as screw connection. The sampler mounting box 13 has a thickness of 10 mm
The accommodating container 16 is attached to the upper surface of the accommodating container 16 and has a heat insulating material 2 made of a foamable elastic material such as neoprene rubber on the outer periphery of the accommodating container 16.
0 is allocated. The length of the collector body 17 is the storage container 1
The portion exposed from 6 has a length that can be gripped by the manipulator 12.

Insulating material 2 provided on the outer periphery of the storage container 16
Regarding 0, the environment of the deep sea floor is generally high water pressure and low temperature, and in order to study microorganisms in this low temperature environment, it is necessary to preserve the sample as much as possible and store it in an experiment. However, with normal heat insulating materials, the heat insulating property cannot be maintained because it shrinks under high pressure.
In this embodiment, the heat insulating material 20 made of a foamable elastic material such as neoprene rubber is used. According to experiments, it has been found that it is effective to wrap neoprene rubber having a thickness of about 25 mm around the storage container 16. Further, even if water is filled in the sampling device mounting box 13 before the storage container 16 is mounted, there is a heat retaining effect, and this may be used together with the heat retaining material 20.

The sample container 19 is, for example, a commercially available container with a lid made of transparent polypropylene capable of autoclave sterilization. The lid is removed and the container is screwed to the lower end of the main body 17 of the sampler to sample the sample. After the sample is collected, the sample container 19 is removed from the sampler main body 17 by closing the lid after moving to the mother ship on the sea and used for the subsequent storage, experiment and research. The material of the sample container 19 is not limited to polypropylene and may be any material that can be sterilized by autoclave, and may be other resin or metal such as stainless steel.

Container with lid The lower end of the sample container 19 has a substantially conical shape, and the conical shape facilitates insertion of the sample container 19 into the seabed. The sample container 19 may be used as it is as a container, or the sample collection port 21 may be opened at the lower end thereof. In addition, FIG.
As shown in, a reinforcing wall 22 may be provided at the tip of the sample container 19. In this case, the sample container 19 may be in the shape of a container or the sample collection port 21 may be formed. Furthermore, FIG.
In the embodiment of FIGS. 2A and 2D, when the sample collection port 21 is formed, a check valve 23 may be provided inside the tip of the sample container 19 as shown in FIG. 2D. Good.

A step portion 25 is formed inside the upper part of the collector main body 17, and a filter 27 sandwiched by two stainless steel screens 26 (about 100 mesh) on the step portion 25.
Is provided. The filter 27 is a bacteria blocking member for preventing bacteria existing in the outside seawater from entering the sample container 19, and is fixed by the lid member 30 via the spacer 29. The lid member 30 hermetically seals the upper portion of the collector body 17, and a pressure equalizing hole 31 is formed inside the lid member 30 to communicate the inside and outside of the collector body 17. The pore size of the filter 27 is 0.2 in order to block passage of bacteria in seawater.
Set to about μm. In addition, as a bacteria blocking member, the filter 2
An elastic member such as rubber is provided instead of 7, and the sample container 19
You may make it equalize the inside and the outside by expansion of an elastic member.

Two O-rings 32 are fixed to the outer circumference of the sampler body 17, and when the sampler body 17 is fitted into the storage container 16, a sealing property between the sample container 19 and the storage container 16 is secured. Then, the sample in the sample container 19 is maintained in a sterile state so as not to be contaminated by external seawater. Also, FIG.
As shown in (B) and FIG. 2 (C), the collector body 17
Sampling ports 33 are formed on both sides or one side of the wall portion. The selection of the sample collection port 21 at the tip of the sample container 19 and the sample collection port 33 at the side is arbitrary, and the sample collection port 21 can be used properly depending on the bottom sediment of the seabed to enable reliable sample collection.

The sampler 15 having the above structure is made of a material capable of autoclave sterilization except for the heat insulating material 20. After the sample container 19 and the filter 27 are set before mounting, the sample collector 15 is sterilized by the autoclave. The heat insulating material 20 is wound, and the sampler mounting box 13 is filled with water and the sampler 15 is mounted.

Next, the operation of the present invention having the above structure will be described. Submersible 1 for taking seabed samples
When a person inside 1 operates the manipulator 12 to grasp the upper part of the sampler body 17 and insert the sampler into a desired position on the seabed, the sample is collected at the sample collection port 2 at the tip of the sample container 19.
1, and depending on the bottom material, when the manipulator 12 moves the collector body 17 in the horizontal direction, the sample enters from the sample collection port 33 on the side of the collector body 17. After the sample is collected, the manipulator 12 fits the sampler body 17 into the storage container 16, and the above operation is repeated for the other sampler 15. When all the samples have been collected, the submersible 11 moves up to the mother ship on the ocean. At this time, in the sample container 19, seawater flows in and out through the filter 27 and the pressure equalizing hole 31 due to changes in pressure and temperature, but bacteria in the seawater from the outside are blocked by the filter 27 and the O-ring 32. Therefore, the inside of the sample container 19 can be maintained in a sterile state, and the microorganisms in the sample container 19 can be prevented from flowing out. Then, the sampler body 17 collected on the mother ship is used for subsequent storage, experiments and research after removing the sample container 19 and closing the lid.

Next, experimental results using the sampler of the present invention will be described. Deep sea submersible "Shinkai 6500"
Equipped with the sampler of the present invention, the water depth of the central axis of the Suruga Trough is 2,625 m, 2,480 m, 2,355 m, 2,22.
Deep seabed mud samples were collected at 4 points of 0 m.

Immediately after collecting the sample, a part of the mud sample was suspended in sterilized filtered seawater, and 0.1 ml of the supernatant was divided into 1 /
It was smeared on Anderson agar medium diluted to 10 concentrations, and this was used for viable cell count. On the other hand, a part of the supernatant is
The solution was heated using a water bath and smeared on an agar medium in the same manner as for viable cells, which was used for spores. Both were cultivated at 25 ° C and 4 ° C for about 1 month, the emerged colonies were counted, and the number of viable cells and the number of spores,
From this, the ratio of spores in the viable cell count was calculated. The culture medium for the viable cell count was further cultured after colony counting, and the emerged colonies were stained with spores to determine whether or not they were spore-forming bacteria, and the ratio of the spore-forming bacteria to the viable cell count was determined. And
The proportion of spores and vegetative cells in the spore-forming bacteria was determined from the number of spores and the proportion of spore-forming bacteria.

As a result, about 70 to 80% of the viable cell count were spore-forming bacteria, and about 80 to 90% of the spore-forming bacteria were present in the form of vegetative cells that were proliferating. Results were obtained. The general idea so far was that spore-forming bacteria were thought to be terrestrial bacteria, but there are many in the deep-sea mud, and it is speculated that many of them are not dormant forms. From this, it was suggested that the presence of marine spore-forming bacteria was suggested.

The present invention is not limited to the above embodiments, and various modifications can be made by those having ordinary knowledge in the technical field to which the present invention belongs. For example, in the above embodiment, an example in which a plurality of storage containers 16 are attached to the collector mounting box 13 of the submersible 11 is shown, but the storage container 16, the collector main body 17, and the sample container 19 are shown.
It is also possible for the diver to collect directly on the seabed using the sample collector 15 consisting of.

[0024]

As is apparent from the above description, according to the present invention, a simple structure including the bacteria blocking member provided in the collector body and the O-ring provided between the collector body and the storage container is used. Since the sterility of the sample can be maintained, it is possible to reduce the size and weight of the sampler. Therefore, when mounting this sampler on a submersible, many sampler can be mounted. Therefore, a large number of samples can be obtained in one dive. Further, since the manipulator simply operates to grip the collector body and move it, the manipulator operation can be simplified. Further, if the sampler mounting box 13 is filled with water or a heat insulating material made of a foamable elastic material is provided on the outer circumference of the storage container, the sample can be stored and used for experiments while maintaining the environment of the deep sea floor. You can

[Brief description of drawings]

FIG. 1 is a perspective view of a sampler of the present invention mounted on a submersible.

2 shows an embodiment of the sampler of the invention, FIG.
2A is a cross-sectional view, FIG. 2B is a cross-sectional view taken along line BB of FIG. 2A in the direction of the arrow, and FIG. 2C is a modification of the sampling port. A cross-sectional view similar to FIG. 2B, FIG.
FIG. 8 is a partial cross-sectional view showing a modified example of the sample container.

FIG. 3 is a schematic cross-sectional view showing a conventional sample collector.

[Explanation of symbols]

11 ... submersible, 12 ... manipulator, 13 ... collector mounting box 15 ... sampler, 16 ... storage container, 17 ... sampler body, 19 ... sample container 20 ... heat insulating material, 21 ... sample collection port, 22 ... reinforcement Wall, 23
... Check valve, 25 ... Step portion 26 ... Screen, 27 ... Filter (bacteria blocking member), 2
9 ... Spacer 30 ... Lid member, 31 ... Pressure equalizing hole, 32 ... O-ring, 33 ...
Sampling port

 ─────────────────────────────────────────────────── ─── Continuation of front page (72) Inventor Yoshiji Imai 2 Natsushima-cho 15 Yokosuka City Kanagawa Prefectural Marine Science and Technology Center (72) Inventor Toshiaki Sakurai 2 Natsushima-cho 15 Yokosuka City Kanagawa Prefectural Marine Science and Technology Center (72) Inventor Shozo Tashiro 2-15 Natsushima-cho, Yokosuka-shi, Kanagawa Prefectural Marine Science and Technology Center (72) Inventor Katsufumi Akazawa 2-15, Natsushima-cho, Yokosuka-shi, Kanagawa (56) Reference Reference JP-A-4-339234 (JP, A) JP-B 1-335158 (JP, B2)

Claims (4)

[Claims] 1. A cylindrical storage container, a cylindrical collector body fitted in the container via an O-ring, and a sample container detachably attached to the lower part of the collector body. A sample collection port formed in at least one of a lower end of the sample container and a side part of the collector body, a bacterium blocking member arranged on an upper part of the collector body, and an upper part of the collector body are closed. A sample collector for collecting a seabed sample in an aseptic state, comprising a lid member and a pressure equalizing hole formed inside the lid member and connecting the inside and outside of the collector body. 2. A submarine sample aseptic according to claim 1, further comprising: a sampler mounting box fixed near the manipulator of the submersible, and a plurality of the storage containers being fixed to the sampler mounting box. Sampling device for collecting in the state. 3. The sample collector for collecting a seabed sample in an aseptic state according to claim 2, wherein water is filled in the sampler mounting box. 4. A sample for aseptically collecting the seabed sample according to any one of claims 1 to 3, wherein a heat insulating material made of a foamable elastic material is provided on the outer periphery of the storage container. Collector.