JPH07175A - Device for culturing nematodes - Google Patents

Abstract

PURPOSE:To provide a nematode-culturing device capable of culturing and producing nematodes in high productivity. CONSTITUTION:The characteristic of this device for culturing nematodes comprises disposing a medium 3 for culturing the nematodes and arranging an active substance 2 (zeolite) for adsorbing waster substances such as ammonia in an air-tight culture space.

Description

Detailed Description of the Invention

[0001]

[Industrial application] Some nematodes enter the nematode's body by invading the insect's blood cavity through the insect's mouth, anus, ankles, etc. while contaminating with the insect. An insect parasitic nematode that releases the symbiotic bacteria that inhabit it, destroys the immune mechanism of the host insect, and proliferates in the body of the host insect, causing the host insect to die is known. There is. Using such insect-parasitic nematodes to kill pests that damage plants such as agricultural crops, in order to prevent damage to the plants, it is considered to culture such nematodes in large quantities. There is.

Therefore, the present invention relates to an apparatus for culturing nematodes which has an airtight culture space and can accommodate a medium for culturing nematodes in the culture space of the apparatus.

[0003]

2. Description of the Related Art Heretofore, this type of nematode culturing apparatus has merely been capable of accommodating a medium in an airtight culturing space.

[0004]

It is generally known that nematodes secrete some kind of waste products when they grow on a medium, so that when they grow, their growth is suppressed. When the nematodes are cultured and proliferated by the conventional nematode culturing apparatus described above, waste products are generated in the culture space, and the culture space is airtight, and thus accumulated in the culture space. Growth is suppressed due to the waste,
Nematode productivity was likely to decrease.

Therefore, in view of the above-mentioned drawbacks, the object of the present invention is to cultivate nematodes with high productivity even if culturing nematodes produces wastes in the culture space and the productivity is not lowered. The purpose of the present invention is to provide a culture device for nematodes that can be produced by

[0006]

A characteristic feature of the present invention for achieving this object is that an adsorptive active substance for waste products of nematodes is arranged in a culture space. Provided with a support for holding a medium for culturing, the support may have an adsorptive active substance, or the support may be formed by mixing an adsorptive active substance, The support may be formed of an adsorptive active substance, a holding part for holding the adsorptive active substance may be formed in the culture space, and further better if the adsorptive active substance is a zeolite. , And its effects are as follows.

[0007]

[Effect] That is, even if nematodes are cultured in an airtight culture space and nematode waste products are generated in the culture space, an adsorption active substance for the waste products is arranged in the culture space. Therefore, since the adsorptive active substance adsorbs the waste products, the waste products do not suppress the growth of the nematodes. Therefore, the productivity of culturing and multiplying nematodes is greatly improved.

[0008] In addition, the medium for culturing nematodes has an adsorbing active substance held on the support, or the adsorbing active substance is mixed with the support, or
If the support is formed of an adsorptive active substance, the adsorptive active substance can be arranged so as not to be bulky in the culture space and to be an obstacle in the culture space.

Further, if the holding portion for holding the adsorptive active substance is formed in the culture space, the adsorptive active substance adsorbs waste products more than its capacity, causing a problem that the adsorptive activity decreases. Even if it is, it can be arranged in the culture space so that the adsorption active substance can be easily exchanged.

Further, if the adsorption active substance is zeolite, it is possible to obtain good adsorption activity for ammonia and the like contained in waste products.

[0011]

EFFECTS OF THE INVENTION Therefore, since it was possible to prevent the growth of nematodes from being suppressed and to greatly improve the productivity of culturing and producing nematodes, for example, Steinernema kusidai. Steinernema Carpocapsae All (Steine)
rnema carpocapsae All) (hereinafter referred to as All),
Sweet potatoes, when cultivated nematodes having the ability to kill pests against agricultural crops such as cabbage, since it can be used as a biopesticide for killing the pests, the pests are killed by the pests. A large amount of nematodes can be provided to the technology that uses nematodes, such that damage to agricultural crops can be greatly reduced and it can be used for increasing crop yields. .

[0012]

Embodiments of the present invention will be described with reference to the drawings. As shown in FIG. 1, a dog food medium 3a is adsorbed and supported on a support A in which a zeolite, which is an example of an adsorption active substance 2, is contained in a polyurethane foam 1, and a surface area and a void are provided between the supports A. In order to secure the above conditions, the supports A were assembled with each other to form a laminate, and sterilization was performed according to a conventional method. When the infested larva of Kushidai was inoculated to this laminate and placed in a closed container B to make a nematode culturing device, culturing was carried out at 25 ° C. for 20 days,
It was found that the proliferative property is improved by about 50% as compared with the case where zeolite is not used.

The dog food medium is dog food ………………………… 8.8g Yeast extract …………………… 1.2g Lard …………………… 4.0g Carboxymethyl cellulose sodium salt: 2.0 g of phosphate buffer solution (1/3 M potassium dihydrogen phosphate (KH ₂ PO ₄ ) and 33 ml of disodium hydrogen phosphate (Na ₂ HPO ₄ ) 4 It was dissolved in 10 times the mixed ratio of 6) and adjusted to 100 ml.

Various culture test examples are shown below.

[Test Example I] Zeolite 2 as an adsorptive active substance is arranged in various forms in the culture space, and nematodes are cultured using dog food medium 3a as medium 3.
The proliferative property was examined.

[Test Example I-1] As shown in FIG.
On a Petri dish 4 having a diameter of mm, a support A having a cubic polyurethane foam 1 of 1 cm square and containing 0.05 g of zeolite 2 having a pore size of 3Å is formed as a surface area and a void between the supports A. And the dog food medium 3a as a whole is
It was adsorbed and supported by 0 ml and sterilized by a conventional method.
This stack was inoculated with 2000 larvae of the infectious agent of Kushidai, and placed in a closed container B to prepare a nematode culture device.
Culture was performed at 5 ° C for 20 days.

[Test Example I-2] As shown in FIG.
On a Petri dish 4 with a diameter of mm, a cubic polyurethane foam 1 of 1 cm square is used as a support A to form a laminate, and 20 ml of the dog food medium 3a is adsorbed and supported as a whole, and further zeolite 2 having a pore diameter of 3Å is supported. In addition to 5 g Petri dish 4, they were mixed and sterilized according to a conventional method. This laminate was inoculated with 2000 larvae of the infectious agent of Kushidai, and cultured in a closed container B at 25 ° C.

[Test Example I-3] As shown in FIG.
On a Petri dish 4 with a diameter of mm, a 1 cm cubic zeolite (30 g) having a pore size of 3 Å was used as a support A to form a laminate, and 20 ml of the dog food medium 3a was adsorbed and supported as a whole, Sterilization was performed according to the method. This laminate was inoculated with 2000 larvae of the infectious agent of Kushidai, and cultured in a closed container B at 25 ° C. (Fig. 3
reference)

[Comparative Example I-1] Petri dish 4 having a diameter of 90 mm
A cubic body of 1 cm square polyurethane foam 1 was used as the support A to form a laminate, and 20 ml of the dog food medium 3a was adsorbed and supported as a whole, and sterilized according to a conventional method. This laminate was inoculated with 2000 larvae of the infectious agent of Kushidai, and cultured in a closed container B at 25 ° C.

[Test Examples I-4 to I-6] Test Examples I-1 to I-6
Incubation was performed in the same manner using Alli infected larva instead of I-3 infected larva.

[Comparative Example I-2] Incubation was carried out in the same manner using an Allel infected larva in place of the Infected larva of Kushidai in Comparative Example I-1.

The results are shown in Table 1.

[0023]

[Table 1]

From Table 1, when Zeolite 2 which is an example of an adsorptive active substance is placed facing the medium 3a in the culture space (Test Examples I-1 to I-6), when Zeolite 2 is not used Compared with (Comparative Examples I-1, I-2), 50-80% for kushidai and 20-30% for All.
It was found that the number of nematodes growing increased. That is, it can be seen that when Zeolite 2 is used, it is possible to provide a nematode culturing device that exhibits high productivity for nematodes.

In addition, the kushidai used in Test Example I,
All is a typical example of nematodes, and nematodes that can be used for culture are not limited thereto. Also, the material used to form the support is not limited to the polyurethane foam 1, and various materials such as foamed resins can be applied.

[Test Example II] Attempts were made to adsorb and remove waste products which suppress the growth property in the gas phase, and the zeolite 2 used as an adsorption active substance was changed to have various pore sizes to improve the growth property. We investigated whether there was an improvement. In this test example, the dog food agar medium 3b was used as the medium 3.

[Test Example II-1] An aluminum cup C was placed in the center of a 150 mm-diameter Petri dish 4 as a holding part for holding an adsorption active substance 2 having a diameter of 50 mm, and 15 g of zeolite 2 having a pore size of 3Å was used as the adsorption active substance. 50 m of dog food agar medium 3b put in aluminum cup C and sterilized under dry heat condition at 150 ° C. for 2 hours and then sterilized according to a conventional method
1 and placed on a petri dish 4 and 10,000 larvae of the infectious agent of Kushidai were inoculated on the medium 3 and cultured at 25 ° C. for 20 days. (See Figure 4)

The dog food agar medium means dog food ………………………… 8.8g yeast extract …………………… 1.2g lard …………………… 4 0.0 g agar …………………………………………………………………………………………………………………………………… 0.3 g is dissolved in phosphate buffer and adjusted to 100 ml.

[Comparative Example II-1] The same test was carried out without placing the zeolite 2 in the aluminum cup C in Test Example II-1.

As a result, in Test Example II-1, Comparative Example
It was found that the number of growth increased by 37% compared to II-1 (see Table 2), and the waste products that suppress the growth of nematodes are present in the air layer and can be removed by adsorption. It was found that, if possible, Zeolite 2 can suppress the decrease in the growth of nematodes and improve the productivity of nematodes even if they are not in contact with medium 3. However, this does not mean that the waste products that suppress the growth of nematodes exist only in the air layer, and do not exclude the possibility that the waste products exist in the medium.

[Test Example II-2] The same test was performed by changing Zeolite 2 in Test Example II-1 to one having a pore size of 4Å.

[Test Example II-3] The same test was performed by changing Zeolite 2 in Test Example II-1 to one having a pore size of 9Å.

The results are shown in Table 2.

[0034]

[Table 2]

As a result, it was confirmed that the proliferative properties were improved by about 30% to 60%, respectively, and it was found that the zeolite 2 used in the nematode culture device having various pore sizes can be used. It was

The aluminum cup C is an example of a holding portion, and may be a plastic cup, a small-diameter petri dish, or the like. Furthermore, the holding portion does not necessarily have to be on the petri dish 4, and waste products in the culture space may be discarded. It suffices if it is at a position where it can adsorb, and a holding part for holding the adsorbing active substance may be provided on the lid of the airtight container that forms the culture space and is opened and closed. (See Figure 5)

[Test Example III] Not only Zeolite 2 but also a substance other than Zeolite 2 was used as the adsorption active substance in Test Example 2
The effectiveness when used in a different arrangement from was investigated.

[Test Example III-1] A petri dish 5 having a diameter of 90 mm was placed at the center of a petri dish 4 having a diameter of 150 mm, and 6 g of zeolite 2 having a pore size of 9 Å was used as an adsorbing active substance in an amount of 150 m.
The dog food agar medium 3b, which is placed on the outside of the 90 mm diameter petri dish 5 on the m diameter petri dish 4 and sterilized under the dry heat condition at 150 ° C. for 2 hours, and then sterilized according to the usual method, is used.
The medium was placed on a petri dish 5 with a diameter of 90 mm and a diameter of 90 mm, and 2000 larvae of the infectious larva of Kushidai were inoculated on the medium and cultured at 25 ° C. for 20 days.

Test Example III-2 A similar test was conducted using powdered activated carbon instead of zeolite 2 having a pore size of 9Å in Test Example III-1.

[Test Example III-3] A similar test was performed using synthetic amorphous silica (Tokushiru P of Tokuyama Soda Co., Ltd.) instead of zeolite 2 having a pore size of 9Å in Test Example III-1.

[Test Example III-4] The same test was conducted using calcium silicate (Florite P manufactured by Tokuyama Soda Co., Ltd.) instead of zeolite 2 having a pore size of 9Å in Test Example III-1.

[Comparative Example III-1] A similar test was conducted by removing Zeolite 2 having a pore size of 9Å in Test Example III-1.

The results are shown in Table 3.

[0044]

[Table 3]

As a result, it was found that the use of activated carbon, silica or calcium silicate other than Zeolite 2 as the adsorption active substance can suppress the decrease in the proliferative ability of nematodes. In addition, in this test example, zeolite 2, activated carbon,
It is needless to say that silica and calcium silicate are examples of the adsorption active substance, and any substance having an adsorption activity for waste products can be used as the adsorption active substance.

From Test Examples I to III, the nematode culturing apparatus of the present invention is provided with a medium for culturing nematodes in an airtight culture space, and the adsorption active substance 2 is added to the culture space. It can be seen that if the structure is arranged inside and the waste products can be adsorbed and removed, the productivity of nematodes can be improved without deteriorating the productivity of nematodes.

It should be noted that reference numerals are added to the claims for convenience of comparison with the drawings, but the present invention is not limited to the configurations of the accompanying drawings by the entry.

[Brief description of drawings]

FIG. 1 is a conceptual diagram showing an example of Test Example I-1.

FIG. 2 is a conceptual diagram showing an example of Test Example I-2.

FIG. 3 is a conceptual diagram showing an example of Test Example I-3.

FIG. 4 is a conceptual diagram showing an example of Test Example II-1.

FIG. 5 is a conceptual diagram showing an example of Test Example II different from Test Example II-1.

FIG. 6 is a conceptual diagram showing an example of Test Example III-1.

[Explanation of symbols]

 A support C holding part 2 adsorbing active substance

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Tadaaki Kawasugi 5-6 Koyodai, Ryugasaki City, Ibaraki Prefecture Tsukuba Laboratory, Kubota Institute for Technology Development

Claims (6)

[Claims] 1. A nematode culturing apparatus comprising an airtight culture space and capable of accommodating a medium for culturing nematodes in the culture space, the adsorption device for waste products of the nematodes. A nematode culturing apparatus in which the active substance 2 is placed in the culturing space. 2. A support A for holding a medium is provided,
The nematode culturing apparatus according to claim 1, wherein the support A holds the adsorptive active substance 2. 3. A support A for holding a medium is provided,
The nematode culturing apparatus according to claim 1, wherein the support A is formed by mixing the adsorptive active substance 2. 4. A support A for holding a medium is provided,
The support A is formed of an adsorptive active substance 2.
The nematode culture device described. 5. The nematode culturing apparatus according to claim 1, wherein a holding section C for holding the adsorptive active substance 2 is formed in the culturing space. 6. The nematode culture device according to claim 1, wherein the adsorptive active substance 2 is zeolite.