Container for sterile feeding of Clania variegata and method for sterile feeding of Clania variegata by using same

Abstract

The present invention addresses the problem of developing and providing a sterile feeding technique for Clania variegata and a sterile feeding container required to implement the technique. The present invention provides a sterile feeding container for Clania variegata, the sterile feeding container comprising a heat-resistant container which is made of a raw material having a heat-resistant temperature of 100 DEG C or more and which is internally in a sterile state, an artificial feed, and a Clania variegata nest material comprising bark, the artificial feed and the Clania variegata nest material being disposed in the container so as to be in contact with each other, and the artificial feed and the Clania variegata nest material being in contact with each other. Alternatively, the artificial feed is disposed at a distance such that the Clania variegata can directly stretch across the Clania variegata nest material and the artificial feed.

Description

Technical Field

[0001] The present invention relates to a sterile breeding container for sedge moths, a method for manufacturing the container, and a sterile breeding method for sedge moths using the container. Background Art

[0002] Bagworms are a general term for the larvae of moths belonging to the family Psychidae in the order Lepidoptera. Bagworm silk, produced by these moths, has recently attracted attention as a new animal-derived natural fiber because it possesses superior physical properties compared to silk produced by silkworms (Patent Document 1 and Non-Patent Document 1).

[0003] The silk industry using moths is still in its infancy, and breeding technology is still under development. Efficient and stable mass breeding technology for moths is essential for the early stages of industrializing moth silk, and establishing this technology is now a top priority.

[0004] Like silkworms, also members of the Lepidoptera order, moths feed on fresh leaves from broadleaf trees. Therefore, collecting and regularly replacing bait leaves is a necessary step in their breeding. However, this process requires considerable time, labor, and cost, becoming a significant obstacle to improving work efficiency and business management.

[0005] A part of the rearing technology of the moth can directly utilize the rearing technology of the silkworm established in the sericulture industry. For silkworms, as a means to solve the above-mentioned problems, a technology (non-patent literature 2) has been established to raise a certain period of time using artificial feed under aseptic conditions. According to this rearing technology, not only can a certain quality of bait be stably supplied, but also feeding management such as bait replacement is not required for a certain period of time. Therefore, if aseptic rearing can also be carried out in the rearing of the moth, the workload can be reduced. However, in the case of the moth, the aseptic rearing technology of the silkworm cannot be directly utilized due to the difference in ecological aspects. One of the differences is the nesting of the moth.

[0006] Unlike silkworms, moths build nests and live within them throughout their larval stages. Nesting begins immediately after hatching, and once built, the nest is rarely replaced. Instead, it is repeatedly repaired and expanded as the moth grows, remaining in use throughout its larval stages. Therefore, nests are essential for moth survival, and the supply of nesting materials and food is crucial in rearing moths.

[0007] The nests of moths are made by winding branches, leaves, etc. with silk spun by moths. Therefore, under normal breeding conditions, moths can use part of the branches and leaves given for feeding as nest materials. However, under sterile breeding, they are fed artificial feed instead of branches and leaves. In this case, moths use artificial feed as nest materials. However, artificial feed tends to become powder due to drying after a few days, resulting in the nest losing strength and soon being damaged. Therefore, there is a problem that moths must use artificial feed as nest materials to rebuild their nests. This problem is particularly serious in young larvae from the first to second instar, which have low resistance to the environment, low walking ability, and little excess energy such as fat.

[0008] Prior art literature

[0009] Patent Literature

[0010] Patent Document 1: WO2019 / 003364

[0011] Non-patent literature

[0012] Non-patent document 1: Osaki Shigeyoshi, 2002, , 58: 74-78

[0013] Non-patent document 2: Ito and Horie, 1962, Journal of Insect Physiology, 8: 569-578 Summary of the Invention

[0014] Problems to be solved by the invention

[0015] The present invention aims to develop and provide a sterile rearing technology for the moth and a sterile rearing container required for realizing the technology.

[0016] Means for solving problems

[0017] As a solution to one of the challenges associated with mass-rearing sedge moths using artificial diets, the inventors developed artificial nesting materials that replace branches and leaves, and a nesting assistance method using these materials (WO2022 / 265120). By using these artificial nesting materials in conjunction with the artificial diet, the aforementioned sterility issue was resolved, successfully establishing a method for sterile rearing of sedge moths.

[0018] In addition, aseptic rearing usually places the sterilized eggs in aseptic rearing containers to raise the larvae that have hatched. As mentioned above, in the case of the brood moth, since nesting begins immediately after hatching, the eggs are preferably arranged on or near the artificial nest material. On the other hand, since the artificial nest material is not food, the larvae cannot be fed during nesting. Therefore, after hatching, the larvae must nest in a fasting state and then seek a place for artificial feed and move. This is a large physical burden for the brood moth just after hatching, and becomes the reason that increases the initial mortality rate of the brood moth.

[0019] The present inventors have conducted extensive research and have discovered that placing the artificial nest material and artificial feed in contact or near each other significantly reduces the distance traveled by hatched moths to build nests and feed, thereby resolving the aforementioned problem. Furthermore, they unexpectedly discovered that placing the artificial nest material on the artificial feed increased the growth of moths.

[0020] The present invention provides the following solutions based on these development results and new findings.

[0021] (1) A sterile breeding container for moths, comprising a heat-resistant container made of a raw material having a heat-resistant temperature of 100°C or above, artificial feed, and moth nest material comprising tree bark, wherein the heat-resistant container is sterile, and within the heat-resistant container, the artificial feed and the moth nest material are arranged in contact with each other, or are arranged at a distance such that moths can directly cross the moth nest material and the artificial feed.

[0022] (2) In the sterile breeding container according to (1), the artificial feed is attached to a portion of the surface of the nest material of the moth.

[0023] (3) The sterile breeding container according to (1) or (2), wherein the heat-resistant container has a vent hole.

[0024] (4) The sterile breeding container according to any one of (1) to (3), wherein the bark is cork.

[0025] (5) The sterile breeding container according to any one of (1) to (4), which is used for first- to fourth-instar worms.

[0026] (6) The sterile breeding container according to any one of (1) to (5), wherein the nest material for the moth is arranged at a position higher than the artificial feed.

[0027] (7) A baited nest material for moths, wherein the artificial feed is attached to a portion of the nest material for moths containing bark.

[0028] (8) The baited nest material for moths according to (7), wherein the bark is cork.

[0029] (9) The baited nest material for moths according to (7) or (8), which is used for moths from the first to the fourth instar.

[0030] (10) The nest material with bait for the moth according to any one of (7) to (9), which is sterilized and packaged.

[0031] (11) A method for manufacturing a sterile breeding container for moths, the method comprising: a configuration step of configuring artificial feed and moth nest materials in a heat-resistant container made of raw materials having a heat-resistant temperature of 100°C or above; a contact step of bringing the artificial feed and moth nest materials into contact in the heat-resistant container; and a sterilization step of sterilizing the heat-resistant container after sealing the interior of the heat-resistant container.

[0032] (12) According to the manufacturing method described in (11), the above-mentioned contact is a state in which the artificial feed of the moth is attached to a part of the surface of the moth nest material.

[0033] (13) The production method according to (11) or (12), wherein the bark is cork.

[0034] (14) According to the manufacturing method described in any one of (11) to (13), the nest material of the moth is arranged at a position higher than the artificial feed.

[0035] (15) A method for aseptic rearing of moth moths, comprising an inoculation step of placing moth moth eggs and / or early-instar moth moths in a sterile rearing container according to any one of (1) to (6) under aseptic conditions.

[0036] (16) The aseptic breeding method according to (15) includes a sterilization step of sterilizing the surface of the eggs of the moth before the inoculation step.

[0037] (17) According to the sterile breeding method described in (16), the above-mentioned sterilization treatment is a contact treatment with 70% ethanol or 2-3% formalin.

[0038] (18) According to the aseptic breeding method described in any one of (15) to (17), the first-instar moth worms are moth worms within 3 days after hatching.

[0039] This description incorporates the disclosure of Japanese Patent Application No. 2023-028377 upon which the present application claims priority.

[0040] Effects of the Invention

[0041] By using the sterile rearing container for moths of the present invention to rear moths aseptically, a large number of moths can be reared without requiring rearing management such as changing bait from the time of hatching to the time of growth to the fourth instar.

[0042] Furthermore, the aseptic rearing method of the present invention can reduce the mortality rate of early-stage moths that are difficult to handle. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 This diagram shows the nesting state of first-instar moths hatched within the sterile rearing container for moths according to the present invention after inoculation. The diagram shows a nest built on the surface of nesting material placed within the sterile rearing container for moths according to the present invention. The conical nest is dotted with cork particles gnawed off the cork mat.

[0044] Figure 2 The graph shows the age composition of the moths in the sterile rearing containers for moths of the present invention 45 days, 90 days, and 105 days after inoculation.

[0045] Figure 3 Schematic diagram of each test area shown in Table 1. In the figure, A shows test area A, B shows test area B, C shows test area C, and D shows test area D. The diagram shows the positional relationship between the nesting material and the artificial feed in the container.

[0046] Figure 4 The figure shows the survival rate of the moth in each test plot 15 days after inoculation. In the figure, A shows test plot A, B shows test plot B, C shows test plot C, and D shows test plot D.

[0047] Figure 5 The average length of the long axis of the nest of the moth in each test plot 15 days after inoculation is shown. In the figure, A shows test plot A, B shows test plot B, C shows test plot C, and D shows test plot D. DETAILED DESCRIPTION

[0048] 1. Sterile breeding container for moths

[0049] 1-1. Overview

[0050] The first embodiment of the present invention is a sterile breeding container for moths (often referred to herein as a "sterile breeding container"). The sterile breeding container of the present invention is characterized by comprising a heat-resistant container, an artificial feed, and moth nest material. Within the heat-resistant container, the artificial feed and moth nest material are arranged so as to be in contact with each other or spaced apart so that moths can directly cross between the moth nest material and the artificial feed. The sterile breeding container of the present invention can store moths for long periods of time without the addition of moths. Furthermore, by introducing moths under sterile conditions, moths can be reared from newly hatched instars to fourth instars without changing the bait.

[0051] 1-2. Definition

[0052] The following terms frequently used in this specification are defined as follows.

[0053] As mentioned above, the term "psychidae" refers to the larvae of moths belonging to the family Psychidae of the order Lepidoptera. Moths of the family Psychidae are distributed worldwide, and the psychidae targeted in this specification may be of any species. For example, although not limited, species such as Acanthopsyche, Anatolopsyche, Bacotia, Bambalina, Canephora, Chalioides, Dahlica, Diplodoma, Eumeta, Eumasia, Kozhantshikovia, Mahasena, Nipponopsyche, Paranarychia, Proutia, Psyche, Pteroma, Siederia, Striglocyrbasia, Taleporia, Theriodopteryx, and Trigonodoma can be cited. Furthermore, the psychidae targeted in this specification are not limited to either sex or age.

[0054] In this specification, the term "growth stage" refers to a criterion for classifying moths based on their growth. In this specification, moths are categorized into three stages: early, middle, and advanced. Moths in each growth stage are referred to as early, mid, and late stages.

[0055] In this specification, "early-stage moths" refer to young moths. Although not limited, moths from the first to second instars that have just hatched and have built nests with a long diameter of 1 mm or more and less than 10 mm are eligible.

[0056] In this specification, the term "mid-stage moth" refers to a moth in its middle stage. Although not limited, moths that build nests with a long diameter of 10 mm or more and less than 20 mm and are approximately 3rd to 4th instar meet this requirement.

[0057] In this specification, the term "late-stage moth" refers to moths at an advanced stage. Although not limited, moths that build nests with a long diameter of 20 mm or more and are approximately 5th instar or older meet this definition.

[0058] In this specification, the term "nest (of the moth)" refers to a bag-shaped movable residence that is shaped like a spindle, cylinder, cone, or the like and wraps the moth's entire body. The nest is made of silk spun by the moth itself and raw materials such as leaves and branches connected by the silk. The moth basically stays in the nest throughout its entire larval stage and does not leave the nest. When feeding or moving, only a part of its body is exposed from the opening of the nest, and it is usually in a state of carrying the nest. The moth does not make a cocoon, and pupation takes place in the nest. As described above, the moth and the nest are in an inseparable relationship.

[0059] In this specification, "nesting" refers to the process by which moths create nests. As mentioned above, moths do not replace the nests they create immediately after hatching, but instead repeatedly repair and expand them as they grow, using them throughout their larval stages. Therefore, nesting in this specification not only refers to building a nest from a pre-existing nest, as occurs immediately after hatching, but also encompasses the broader concept of nesting, which includes repairing or expanding existing nests to optimize their growth. "Repairing and expanding a nest" refers to expanding the nest or repairing damaged parts to suit the moth's growth and the moth's condition. Nest repair and expansion are typically accomplished by adding new nesting materials or reinforcing moth silk.

[0060] As used herein, "silk" refers to silk produced from insect protein, spun by insect larvae and adults for purposes such as nest building, locomotion, anchoring, cocoon making, and bait capture. Furthermore, "silk from the ground moth" as used herein refers to silk spun by the ground moth.

[0061] In this specification, "sterilization" means reducing the survival probability of bacteria and viruses coexisting in an object to 1 in 1,000,000 or less, preferably 0. "Sterilization treatment" means treatment to sterilize an object.

[0062] As used herein, "sterilization" refers to significantly reducing the number of bacteria adhering to or coexisting with an object, thereby inactivating their infectivity. "Bactericidal treatment" refers to a treatment that sterilizes an object. Similar to sterilization, this treatment essentially kills bacteria without restriction, but the treatment conditions are milder than sterilization, and the bacterial destruction is less stringent than in sterilization.

[0063] The term "sterile state" as used herein refers to a state in which no bacteria exist, and is particularly used herein to refer to a state in which no bacteria exist in a closed space.

[0064] In this specification, the term "sterile breeding" refers to breeding the target organism in a sterile state. In this specification, unless otherwise specified, it means breeding the moth under a sterile state.

[0065] In this specification, the term "aseptic conditions" refers to conditions that can satisfy the aseptic state. The term "under aseptic conditions" refers to an environment that can satisfy the aseptic state.

[0066] 1-3. Composition

[0067] In this specification, a "sterile breeding container for moths" (often referred to simply as a "sterile breeding container") refers to a breeding container for moths whose interior is maintained in a sterile state. The growth stage of moths to be the target of the sterile breeding container in this specification is not limited. However, due to the possibility that the amount of food increases when the moths reach the sub-terminal or final stages, the artificial feed pre-placed in the container may become insufficient, the possibility of artificial feed deterioration due to long-term storage at room temperature, and the hygienic problems caused by the increase in molting shells and feces of the moths as they grow, it is generally preferred to use it for moths in the early to middle stages, that is, from newly hatched first-instar to fourth-instar stages.

[0068] 1-3-1. Components

[0069] The sterile breeding container of the present invention comprises a heat-resistant container, artificial feed, and a nest material for the moth as essential components, and further comprises a ventilation hole as an optional component.

[0070] (1) Heat-resistant container

[0071] The so-called "heat-resistant container" is the shell of the sterile breeding container of the present invention, and refers to a container capable of maintaining a sterile state inside.

[0072] The material of the heat-resistant container is not limited as long as it has a heat resistance of 100°C or higher and is impermeable to bacteria. A heat resistance of 100°C or higher is required because the heat-resistant container must be sterilized during the production of the sterile breeding container of the present invention. Specific examples of the material include synthetic resins, glass, metals, silicon, ceramics, stone, plant-based materials (e.g., paper), animal-based materials (e.g., chrome-tanned cowhide), or combinations thereof. "Synthetic resins" herein include plastics (thermoplastic resins), thermosetting resins, and synthetic rubbers that have a heat resistance of 100°C or higher, 110°C or higher, and preferably 120°C or higher. Plastics with this heat resistance are not limited; examples include polyester (including polyethylene terephthalate), polyethylene, polypropylene, polyvinyl alcohol, nylon, Teflon (PTFE: a registered trademark), polycarbonate, methylpentene, and phenolic resins. Glass includes quartz glass, fused silica, synthetic quartz, alumina, sapphire glass, ceramics, forsterite, and photosensitive glass. Metals include pure metals such as gold, silver, copper, aluminum, tungsten, molybdenum, chromium, platinum, titanium, and nickel, as well as alloys such as stainless steel, Hastelloy, Inconel, Monel, and Duralumin. Heat-resistant containers can be made from a single material or a combination of two or more.

[0073] In order to confirm the internal state of the sterile breeding container and the growth of the moth during sterile breeding, at least a portion of the heat-resistant container is preferably made of a light-transmitting material that allows visual inspection of the interior of the container. Examples include glass and the above-mentioned plastics (polyester, polypropylene, polyvinyl alcohol, nylon, Teflon, polycarbonate, methylpentene, and phenolic resin).

[0074] The shape of the heat-resistant container is not particularly limited. Generally, any container having a box-like shape (including a cubical or cylindrical shape) with sufficient rigidity to maintain a fixed shape and a space with a fixed internal shape will suffice. It may also be a capsule-like shape (including a spherical shape), a bag-like shape such as an autoclave bag, or a combination thereof. Specific examples include polypropylene food storage containers.

[0075] The capacity of the heat-resistant container can be appropriately determined based on the number of individuals of the moths to be reared in the sterile rearing container of the present invention and the rearing period. For example, when rearing larvae of the large-combed moth (Lycodon spp.) from hatching to the fourth instar, if the number of larvae is 100 or less, the capacity can be 200 mL or more, 300 mL or more, 400 mL or more, or 500 mL or more.

[0076] The heat-resistant container may have an opening and a lid.

[0077] The "opening" is a hole used to insert the artificial feed and moth nest materials described below into the heat-resistant container during the manufacture of the sterile breeding container of the present invention. It can also be used as an insertion hole for moths during sterile breeding of moths. The size of the opening is not limited as long as it is large enough to insert the artificial feed and moth nest materials. A single opening is sufficient in the sterile breeding container of the present invention, but multiple openings are also possible.

[0078] The "lid" is a portion used to close the opening. Therefore, the sterile breeding container of the present invention has the same number of lids as openings. As long as the lid is configured to maintain the sterility of the sterile breeding container by closing the opening, its shape, size, and material are not particularly limited. The connection between the opening and the lid can be either an airtight, tight fit or a breathable, interlocking fit. For example, in the case of a box-shaped heat-resistant container with an opening located on its top surface, the lid can be a cover-type structure, covering the opening from above like a lid. In this case, the connection between the two is an airtight, interlocking fit. Alternatively, the lid can be a screw-on structure, screwed into the heat-resistant container's opening like a nut, or a press-on structure, pressed into the heat-resistant container's opening like a plug. In these cases, the connection between the two is an airtight, tight fit. These lids can be openable and closable, integrated with the heat-resistant container, or detachable. Alternatively, if the heat-resistant container is bag-shaped, the lid may be zipper-shaped. Furthermore, the lid may be formed by pressing or welding the opening. In these cases, the joint between the two also becomes an airtight seal.

[0079] (2) Artificial feed

[0080] Artificial feed is an essential component of the sterile breeding container of the present invention.

[0081] In this specification, "artificial feed" refers to artificially prepared food for moths. In the wild, moths primarily feed on fresh leaves of broadleaf trees. However, in principle, natural leaf bait cannot be provided in sterile breeding containers where bait is not changed. Therefore, artificial feeds are used as an alternative to leaf bait, as they are similar to natural leaf bait, to encourage moths to consume and absorb the necessary water and nutrients for growth.

[0082] The artificial feed used in the sterile breeding container of the present invention is not limited in composition and shape as long as it has the function of serving as the above-mentioned alternative bait, can maintain the quality required for high temperature treatment and long-term use, and is solid (including semi-solid forms such as gel and paste). For example, in addition to the known artificial feed for silkworms (Pingshan, 2020, 89(2): 91-96), commercially available artificial feeds such as Insecta (Nippon Nosan Kogyo Co., Ltd.) and SilkMate (Nippon Nosan Kogyo Co., Ltd.) can also be used.

[0083] The weight of the artificial feed configured in the sterile breeding container of the present invention can be appropriately determined based on the number of individuals of the moths raised in the container and their breeding period. For example, the larvae of the large-nested moth (L. spp.) consume approximately 0.1g to 0.5g of Insecta LFM per larva from the time of hatching to the 4th instar. Therefore, when 10 L. spp. are raised in a sterile breeding container from the time of hatching to the 4th instar, more than 5g of the above-mentioned Insecta can be pre-configured. On the other hand, the artificial feed can also be added from the opening as needed during the sterile breeding period. In this case, the artificial feed that has been sterilized in advance can be added to the sterile breeding container under sterile conditions.

[0084] (3) Materials for moth nests

[0085] The nest material of the moth is an essential component of the sterile breeding container of the present invention.

[0086] In this specification, "nest material for moths" (often referred to simply as "nest material") refers to the components of moth nests and refers to the materials introduced from outside when building their nests. Moths typically wrap plant leaves and twigs around the surface of their nests for purposes such as color protection and reinforcement. In nature, these leaves and twigs serve as nest material.

[0087] (Raw materials for nesting materials)

[0088] The raw materials used in the nest materials of the present invention include all materials artificially supplied in captive environments and used by the moth as nest materials. Examples include tree bark, wood, paper, resin, fiber, and hair. Among them, tree bark is the essential and primary component (raw material) of the nest materials of the moth as used herein.

[0089] The term "bark" refers to a cork tissue consisting of a cork layer located on the outer side of the cork cambium in the trunk, branches, and roots of woody plants. The term "woody plant" refers to a perennial plant with a lignified cambium. The term "tree" refers to a tree. The cork layer is composed of dead cork cells. The species of tree that is the source of the bark is not limited, but is preferably a dicot or gymnosperm. There is no limit to their species. If it is a dicot, for example, plants belonging to the orders Fagales, Juglandales, Hamamelidales, Laurales, Ebenales, Salicales, Ericales, Rosales, and Sapindales can be cited. In addition, if it is a gymnosperm, for example, plants belonging to the Coniferales, Taxales and Cycadales, etc., and Ginkgo biloba can be mentioned. The bark can be the bark of a single tree species or a mixed bark of multiple species. The bark of Fagaceae plants and Pinaceae plants with thick cork layers is suitable. Among them, the bark of Quercus plants and Pinus plants is more preferred as the bark in this specification. Particularly preferred is the bark of European cork oak (Q. suber) and cork oak (Q. variabilis), which are easy to obtain, have relatively stable quality, and are inexpensive. The so-called "cork" is the unprocessed bark or processed bark of European cork oak.

[0090] In this specification, bark includes both processed and unprocessed bark. "Processed bark" refers to bark that has been collected and then pulverized, hollowed out, or sliced, and then shaped as needed. Examples include cork powder obtained by pulverizing cork made from the bark of European Quercus variabilis (Quercus variabilis) into granular form, cork chips pulverized into small flakes, cork sheets compressed from cork powder or cork chips into plate-like shapes, cork mats, hollowed cork, and bark chips pulverized into blocks.

[0091] (shape / size of nest material)

[0092] The shape and size of the nesting material arranged in the sterile breeding container of the present invention are not particularly limited. However, the research results of the present inventors have made it clear that the shape and size of the nesting material preferred by the moth moth vary depending on the growth stage of the moth moth (WO2022 / 265120). Since the sterile breeding container of the present invention is used for moth moths from the first instar to the fourth instar, it is preferably a shape and size suitable for moth moths from the early to mid-stage. In particular, early stage moth moths are generally weaker and have a high mortality rate, so it is desirable to use a nesting material with a shape and size suitable for early stage moth moths.

[0093] In their early stages, the moth is not restricted to nesting materials of any shape, and can readily utilize materials ranging from granules to plates and blocks. However, in their early stages, due to their small mouthparts and weak bite force, they struggle to utilize hard wood and tough fiber as nesting materials. Therefore, relatively soft, easily scraped bark, such as the corkwood of dicots and gymnosperms, is suitable.

[0094] On the other hand, mid-stage moths have preferences for the shape and size of nest materials. The most suitable shape for mid-stage moths is granular nest materials with a specific size.

[0095] The so-called "granular nesting material" refers to a nesting material in the form of particles. The shape of the particles is not particularly limited and includes all shapes such as spherical, approximately spherical, square, modern and ancient square, and amorphous. The shape and size of each particle can be uniform or different. As a specific example, bark crushed into powder or granules is suitable. If it is such a granular nesting material, the mid-stage moth does not process the supplied nesting material, but uses it in its original state or by slightly processing it such as cutting off a part of it. As a result, the labor of the moth in building the nest is greatly reduced, and the nesting rate can be significantly increased.

[0096] The size of the granular nesting material can be any size as long as its major diameter is within the range of 0.3 mm to less than 5.0 mm, 0.5 mm to 4.5 mm, 0.8 mm to 4.0 mm, 1.0 mm to 3.5 mm, or 1.5 mm to 3.0 mm. Granular nesting material with a major diameter of approximately 3.0 mm, i.e., 2.0 mm to 4.0 mm or 2.5 mm to 3.5 mm, is particularly preferred as nesting material for mid-stage moths because it not only increases the nesting rate but also significantly accelerates the growth and development of moths. In this specification, the term "major diameter" refers to the width of the nest material connecting the longest ends. For example, if the nesting material is substantially spherical, its diameter is equivalent to 1.5 mm.

[0097] In addition, in addition to granular nest materials, mid-stage moths can also use plate-shaped nest materials. The so-called "plate-shaped nest material" refers to a relatively thin sheet-shaped nest material. The sheet area is not limited, but it is preferably longer than the body length of the moth in both vertical and horizontal directions. As described later, since moths are chopped into the best shape, the sheet shape of the nest material provided is not limited. It can be square, approximately square, round, approximately round, polygonal, approximately polygonal, and amorphous. In addition, the thickness is not limited and can be 0.2mm to 10mm, 0.5mm to 8mm, 0.8mm to 8mm, 1mm to 6mm, 2mm to 5mm, or 3mm to 4mm. As for the plate-shaped nest material, a thin bark layer obtained by slicing bark, a cork sheet obtained by pressing bark particles obtained by crushing bark into sheets, etc. are suitable.

[0098] Taking the above into consideration, granular nest materials are suitable for early to mid-stage sedge moths. On the other hand, granular nest materials have the side effect of being difficult to operate and manage when placing the nest material in contact with the artificial feed and placing it at a position higher than the artificial feed in the artificial feed configuration in the heat-resistant container described later. On the other hand, plate-shaped nest materials can also be suitably used for early to mid-stage sedge moths as described above. If the nest material is plate-shaped, there is no difficulty in operation and management in the configuration described above. In addition, a three-dimensional configuration such as a vertical plate, box, cylinder or cone can be performed, so the limited space in the sterile breeding container can also be utilized. Therefore, the nest material used in the sterile breeding container of the present invention is more preferably a plate-shaped nest material. Nest materials of different shapes can also be used in combination. For example, a combination of a plate-shaped nest material and a granular nest material can be mentioned.

[0099] As mentioned above, as the moths grow, they repair and expand their nests, so nest materials also need to be provided frequently. The weight of the nest materials placed in the sterile breeding container can be appropriately determined according to the capacity of the container, the number of moths being raised, and the breeding period. For example, from the time of hatching to the 4th instar, each larva of the large-nest moth requires about 0.01g to 0.1g of nest material (cork). Therefore, when 10 larvae of the large-nest moth are raised in a sterile breeding container from the time of hatching to the 4th instar, more than 1g of nest material can be pre-placed. On the other hand, nest materials can also be added from the opening as needed during the sterile breeding period. In this case, the nest materials that have been sterilized in advance can be added to the sterile breeding container under sterile conditions.

[0100] (4) Ventilation holes

[0101] The "vent hole" is an optional component of the sterile breeding container of the present invention, and is a hole provided in the heat-resistant container in order to provide air permeability inside the sterile breeding container of the present invention.

[0102] In addition to being an exhaust hole for balancing the internal pressure and external pressure in the sealed heat-resistant container during sterilization treatment in an autoclave, etc. during the manufacture of the sterile breeding container of the present invention, the vent hole also serves as a gas exchange hole inside the container during the sterile breeding of the moth and a moisture-dehumidification hole to prevent high humidity in the container.

[0103] The vent hole is preferably not a simple hole, but is constituted in a manner that can maintain the aseptic state inside the aseptic breeding container. For example, the vent hole can be opened to the container outside via a sterile filter.

[0104] The so-called "sterile filter" refers to a filter that is air-permeable and blocks the invasion of pollutants. Therefore, pollutants outside the sterile breeding container cannot invade the inside of the container through the vents. In this manual, the so-called "pollutants" refer to spores of microorganisms, filamentous fungi, and plant pollen that are the cause of contamination. There is no limitation on the sterile filter, and a membrane filter with a pore size of 0.2 μm or less can be used. In addition to the above-mentioned properties, the raw materials of the film are not limited as long as they are heat-resistant to 100°C used in steam sterilization. Asymmetric polyethersulfone (aPES) film, cellulose acetate (CA) film, nylon film, polytetrafluoroethylene (PTFE) film, cellulose nitrate (CN) film, glass filter film, etc. can be used. Commercially available sterilization filters can be used.

[0105] The shape and size of the vent hole are not limited. Any shape and size that maintains the desired air permeability and moisture-wicking properties of the container interior is sufficient. However, if one vent hole is too large, the rigidity and strength of the sterile breeding container may be reduced. Therefore, the area of ​​one vent hole is preferably less than 1 / 20, less than 1 / 30, less than 1 / 40, less than 1 / 50, less than 1 / 60, less than 1 / 70, less than 1 / 80, less than 1 / 90, or less than 1 / 100 of the surface area of ​​the entire container.

[0106] In order to improve the air permeability and moisture removal in the container, a plurality of vent holes having an area of ​​1 / 1000 or less relative to the surface area of ​​the entire container may be provided in the sterile breeding container.

[0107] 1-3-2. Configuration

[0108] The sterile breeding container of the present invention is characterized in that the artificial feed is arranged in the heat-resistant container in a manner that the artificial feed is in contact with the nest material of the moth, or at a distance that allows the moths to directly cross the nest material and the artificial feed.

[0109] In the case of contact, the artificial feed placed in the sterile breeding container only needs to be in contact with at least a portion of the nesting material. For example, only a portion of the nesting material may be in contact with the artificial feed, or a portion of two or more separate nesting materials may be in contact with the artificial feed. Specifically, in the case of a plate-shaped nesting material, only one side may be in contact with the artificial feed, or only the lower halves of both sides may be in contact with the artificial feed. Furthermore, in the case of a combination of granular and plate-shaped nesting materials, only a portion of the plate-shaped nesting material may be in contact with the artificial feed, while the granular nesting material may not be in contact.

[0110] The contact method is not limited. It can be achieved by pressing separate solid artificial feeds against the nesting material, or by applying a paste of artificial feed to a portion of the surface of the plate-like nesting material. Alternatively, it can be achieved by embedding a portion of the nesting material in a semi-solid artificial feed, or by mixing multiple granular artificial feeds with multiple granular nesting materials. Alternatively, a combination of these can be used.

[0111] The artificial feed and the moth nest material are preferably arranged in such a way that the nest material is higher than the artificial feed relative to the vertical axis. As shown in the examples described below, it is clear that moth growth is promoted in this arrangement compared to other arrangements.

[0112] Furthermore, when the artificial feed is separated from the nesting materials of the moth, this distance is the distance that the moth can directly cross between the artificial feed and the nesting materials. The phrase "able to directly cross" here means that the moth can move directly between the two without passing through any other places besides the artificial feed and the nesting materials, such as by stretching the insect body. Therefore, the distance that can be directly crossed varies depending on the species and body length of the moth being raised. Although not limited, as specific examples of the distance of this gap, for early-stage moths of the large-nest moth, it can be 1.0mm to 10.0mm, for mid-stage moths, it can be 1.0mm to 15.0mm, for early-stage moths of the tea moth, it can be 0.5mm to 5.0mm, and for mid-stage moths, it can be 0.5mm to 10.0mm.

[0113] There are no specific restrictions on the placement of the artificial feed and the nesting materials for the moth within the heat-resistant container. If they are in close contact, movement of the moth within the container will be hindered, causing unnecessary stress to the moth. Therefore, it is preferable to leave a gap of at least 1 cm, at least 1.5 cm, or at least 2 cm between them, excluding the contact area. For example, a configuration that simulates the natural habitat of moths is suitable. Specifically, the nesting materials can be evenly arranged, for example, by spreading branches of wood within the container with appropriate spacing.

[0114] The positions of the artificial feed and the nesting material of the moth in the heat-resistant container are not particularly limited. For example, they can be arranged at the bottom of the heat-resistant container, or the nesting material can be fixedly arranged at the upper part relative to the vertical axis in the heat-resistant container, that is, at the top of the container. In the latter case, it is more preferable to arrange the artificial feed below the fixed nesting material. Since moths have the habit of moving toward higher positions, if the nesting material is fixed at the top, the moths will live around the nesting material and artificial feed arranged at a high position. In this case, since the excrement falls to the bottom of the container, the moths can be separated from the feces, which is also preferable in terms of hygiene and the health of the moths.

[0115] The location where the vent hole is arranged is not particularly limited, but in consideration of functions such as air permeability and moisture discharge, the vent hole may be arranged on the upper portion of the heat-resistant container or in a position close thereto.

[0116] 1-4. Effect

[0117] By using the sterile breeding container for the moth of the present invention, sterile breeding can be performed without the need for food replacement and can prevent infection caused by bacteria, etc. As a result, the labor, working time and breeding costs in mass breeding of moths can be greatly reduced.

[0118] Furthermore, by using the sterile rearing container for bagworms of the present invention, the survival rate of early-stage bagworms, which have low resistance to environmental changes and physical stimulation and are particularly difficult to handle, can be increased.

[0119] Furthermore, by using the sterile rearing container for bagworms of the present invention, the growth of bagworms can be promoted.

[0120] 2. Moth nest materials with bait

[0121] 2-1. Overview

[0122] The second embodiment of the present invention is a baited nest material for moths. This nest material is characterized by having artificial feed attached to a portion of the nest material. Using this baited nest material for moths can improve the survival rate and growth rate of early-stage and mid-stage moths, which have low environmental resistance and mobility.

[0123] 2-2. Structure

[0124] The baited nest material for moths of the present invention comprises moth nest material and artificial feed as essential components. The basic composition of the moth nest material and artificial feed is the same as that of the moth nest material and artificial feed constituting the sterile breeding container for moths of the first embodiment.

[0125] Furthermore, the configuration in which the artificial feed is attached to a portion of the nest material of the moth moth also follows the contact between the nest material of the moth moth and the artificial feed in the sterile breeding container for moth moths according to the first embodiment. For example, in the case of a plate-shaped nest material, the artificial feed may be attached to only one side or to the lower half of both sides.

[0126] The attachment method is not limited. The artificial feed can be attached to the nesting material by pressing a separate solid artificial feed against the nesting material, or by applying a paste-like artificial feed to a portion of the surface of the plate-like nesting material. Alternatively, the nesting material can be provided with a recessed portion and filled with artificial feed, or a semi-solid artificial feed can be buried in a portion of the nesting material.

[0127] Furthermore, the baited moth nest material can be sterilized and packaged as needed. For example, the baited moth nest material with the above-described structure can be packaged in a sterilization bag, sterilized using an autoclave or gas sterilization, and then sealed. The interior of the packaging container can be degassed. Each feeding amount can also be individually packaged. This form of baited moth nest material facilitates long-term storage and transportation, and can be opened and used immediately when necessary, which is convenient.

[0128] Furthermore, when the baited nest material of the present invention is placed in a breeding container, it is preferably positioned higher than the artificial feed relative to the vertical axis, although this is not a limitation. This is because, as mentioned above, the growth of the moth is promoted in this arrangement compared to other arrangements.

[0129] 2-3. Effect

[0130] Young moths are very difficult to handle due to their low resistance to environmental changes and physical stimulation. Even when fed fresh, natural leaves, their survival rate is only about 30%. This low survival rate is a major problem when raising large numbers of moths for generations. However, the baited moth nesting material of the present invention can increase the survival rate to approximately 100% from 33 to 35 days after hatching.

[0131] 3. Method for manufacturing a sterile breeding container for moth moths

[0132] 3-1. Overview

[0133] The third aspect of the present invention is a method for producing the sterile rearing container for moth moths according to the first aspect. The production method of the present invention can be achieved by placing moth moth nest materials and artificial feed in a heat-resistant container, sealing the opening, and performing a sterilization treatment.

[0134] 3-2. Methods

[0135] The production method of the present invention includes an arrangement step, a contact step, and a sterilization step as essential steps. Each step will be described below.

[0136] 3-2-1. Configuration process

[0137] The "arrangement step" involves placing artificial feed and moth nest materials within a heat-resistant container. This step aims to pre-arrange nest materials and food necessary for moth rearing before sterilizing the interior of the sterile rearing container. The heat-resistant container, artificial feed, and moth nest materials used in this step are identical to the components of the sterile rearing container for moths described in the first embodiment, so a detailed description will be omitted here.

[0138] The capacity of the heat-resistant container is determined in consideration of the number of moths in the sterile rearing container and the rearing period. For example, when rearing 100 or fewer moths from hatching to 4th instar, as described above, the capacity is at least 200 mL, preferably 400 mL or more.

[0139] Artificial feed and nest materials of bagworm moth are inserted into the interior of the container from the opening of the heat-resistant container. At this time, it is desirable to pre-arrange the nest materials and artificial feed in the prescribed configuration when aseptic breeding is completed. For example, the nest materials can be brought into contact with the artificial feed, and further, at this time, it is preferably arranged in a manner that the nest materials are located above the artificial feed. In addition, the weight of the nest materials and artificial feed can also be determined by considering the number of bagworm moths in the manufactured aseptic breeding container and the breeding period. As an example, when raising less than 100 bagworm moths from hatching to the 4th instar, the cork of the nest materials can be at least 1g, 2g, 3g, 4g or 5g, preferably at least 10g, 15g, 20g, 25g or 30g, and the artificial feed can be at least 10g, 15g, 20g or 25g, preferably at least 30g, 35g, 40g, 45g or 50g.

[0140] During the placement process, the artificial feed, the nest material of the moth, and the interior of the heat-resistant container may all be in an unsterilized state. This process is not limited, but is preferably performed in a room with little dust in the air and no wind.

[0141] 3-2-2. Contact process

[0142] The "contact step" involves bringing the artificial feed into contact with the moth nest material within a heat-resistant container. This step aims to improve the survival rate of early- to mid-stage moths during sterile rearing and to create an environment for efficient moth growth under sterile conditions.

[0143] The contact method and arrangement direction of the artificial feed and nesting materials were in accordance with the method described in "1-3-2. Arrangement" of the sterile breeding container for moths described in the first embodiment.

[0144] As long as the artificial feed and the nest material of the moth are in contact with each other in the heat-resistant container, the contact period is not limited. Therefore, the order of the above-mentioned configuration process and this process is not limited. It can be any of before the configuration process, during the configuration process, or after the configuration process. For example, the artificial feed and the nest material can be brought into contact with each other before the configuration process, and then directly configured in the heat-resistant container in this state, or the artificial feed and the nest material can be brought into contact with each other when they are configured in the heat-resistant container. Alternatively, in the case where the heat-resistant container is in the shape of a bag, the artificial feed and the nest material can be placed in the container and the opening can be closed with a lid, and then they can be operated from the outside of the heat-resistant container to bring them into contact.

[0145] 3-2-3. Sterilization process

[0146] The "sterilization step" is a step of sterilizing the heat-resistant container after sealing the interior of the heat-resistant container. This step is performed after the placement step to sterilize the interior of the heat-resistant container.

[0147] In this step, "sealing the interior of the heat-resistant container" means sealing the opening of the heat-resistant container with a lid. This operation creates a sealed space within the heat-resistant container containing the artificial feed and nesting materials. By performing the sterilization treatment in this state, the interior of the heat-resistant container remains sterile.

[0148] The so-called "sterilization treatment" refers to treating an object by a sterilization method. Regarding sterilization methods, various methods are known in this field. For example, high-pressure steam sterilization (autoclave method), boiling sterilization, steam sterilization, ethylene oxide gas sterilization, gamma ray sterilization, dry heat sterilization, etc. can be mentioned. However, the sterile breeding container for the moth of the present invention has a complex internal shape with artificial feed and nest materials arranged inside the container. In addition, the artificial feed is water-soluble and has the possibility of deterioration of quality due to oxidation and thermal drying. Therefore, although not limited, the preferred sterilization method in this process is high-pressure steam sterilization or steam sterilization.

[0149] High-pressure steam sterilization can be carried out according to conventional methods. For example, the heat-resistant container can be placed directly or in a sterilization bag in the pressure chamber described above and sterilized at 2 atmospheres at 115°C for 30 minutes, 121°C for 20 minutes, or 126°C for 15 minutes.

[0150] Steam sterilization is a sterilization method that utilizes steam, similar to high-pressure steam sterilization, but differs in that it is performed at 100°C under atmospheric pressure (1 atm). The basic operation of steam sterilization can also be performed according to conventional methods. For example, a method can be used in which steam treatment is performed at 100°C for 30 minutes, 40 minutes, 50 minutes, or 1 hour, followed by retorting for 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 1 hour, as needed.

[0151] In high-pressure steam sterilization and steam sterilization, if the heat-resistant container does not have a vent hole and the interior of the container is airtight, it is desirable to slightly loosen the joint between the opening and the lid beforehand to equalize the air pressure inside and outside the container. For example, if the lid is a screw cap, the lid can be slightly loosened.

[0152] 4. Aseptic breeding method of moth

[0153] 4-1. Overview

[0154] The fourth aspect of the present invention is a method for aseptically rearing moths. This aseptic rearing method utilizes the aseptic rearing container for moths described in the first aspect to aseptically rear moths. The aseptic rearing method of the present invention allows for the rearing of large numbers of moths, including those in the early to mid-stages of newly hatched moths, which are difficult to handle, without requiring specialized rearing management. This significantly reduces the labor, working time, and rearing costs associated with rearing moths.

[0155] 4-2. Methods

[0156] The aseptic rearing method of the present invention is based on the premise of using the aseptic rearing container for moth moths described in the first embodiment, placing moth moth eggs (often referred to as "eggs" in this specification) and / or first-instar moth moths in the container, and rearing the moth moths from the time of hatching or the first instar.

[0157] The sterile rearing method of the present invention includes an inoculation step as an essential step and, when the inoculation target in the sterile rearing container for the moth is eggs, a sterilization step as an optional step.

[0158] 4-2-1. Sterilization process

[0159] The "sterilization step" is a step of sterilizing the surface of eggs used in the sterile rearing of the moth of the present invention. This step is intended to eliminate contaminants that have passed through unhatched eggs in the sterile rearing container for moths.

[0160] In the aseptic rearing method of the present invention, if the inoculated eggs do not hatch, there is a possibility that filamentous fungi, etc., will be generated through the unhatched eggs, causing contamination within the container. Since the contaminants that cause this are attached to the surface of the eggs, they can be easily removed by sterilizing the egg surface before the inoculation step.

[0161] The method for obtaining the eggs used in this process is not limited. Female moths of relatively new species in the family of Gnaphalidae, such as the large-nest moth and the tea moth, pupate, emerge and mate in a nest after the larval stage, and lay eggs in the pupal shell inside the nest. Therefore, eggs can be obtained from the nest entered by the female moth as a mass of about 1,000 to 3,000 eggs. However, it is difficult to confirm the state of the inside of the nest from the outside, so it is not easy to obtain eggs. Therefore, in order to obtain eggs more efficiently, they can be obtained based on the egg collection method described in Japanese Patent Application Laid-Open No. 2022-092616 developed by the inventors. This method uses a moth egg collector composed of a tubular container having an insertion hole with an inner diameter in which a female moth adult moth fits, a receiving portion capable of accommodating all or part of the abdomen of the female adult moth, and an egg-retaining portion for holding the eggs after laying. After placing a female adult moth separated from its nest and pupal case in the ovipositor, mating is performed with a male adult moth, and then the egg masses laid in the egg-retaining portion are recovered. Using an ovipositor with an egg-retaining portion made of a transparent or light-transmitting material makes it easy to confirm whether eggs have been laid, making it easier to retrieve the eggs.

[0162] The time period for sterilizing eggs is not limited. They can be eggs immediately after spawning or eggs immediately before hatching. However, eggs immediately before hatching have low resistance to external physical stimulation and a high mortality rate. Therefore, eggs that have developed to a certain extent after spawning are preferably sterilized. For example, eggs several days before hatching are suitable.

[0163] The eggs, either in a mass or individually as needed, are then surface sterilized. The sterilization method is not limited, as long as it can eliminate contaminants such as bacteria adhering to the surface without affecting the eggs themselves. Known sterilization methods can be used. Examples include contact with ethanol or formalin. More specifically, for example, the eggs can be contacted with 70% ethanol or 2-3% formalin for 20 to 60 seconds, 30 to 50 seconds, or 35 to 40 seconds. "Contact" here means immersion, spraying, coating, or a combination thereof. Immersion is preferred.

[0164] After this step, the eggs can be washed with sterile water once or several times. In addition, in order to prevent contaminants from re-attaching to the surface of the eggs after the sterilization step, it is desirable to perform this step under sterile conditions such as in a clean bench.

[0165] 4-2-2. Inoculation process

[0166] The "inoculation step" is a step of placing eggs and / or first-instar moths under aseptic conditions in the sterile rearing container for moths described in the first embodiment. In this specification, placing eggs and / or first-instar moths in the sterile rearing container for moths is referred to as "inoculation."

[0167] To prevent contaminants from entering the sterile culture container, inoculation is performed under sterile conditions. There are no specific sterile conditions, and inoculation can usually be performed in a clean bench or other similar environment.

[0168] The basic operation of this step can be applied to inoculation techniques using Escherichia coli, cultured cells, and the like. For example, under sterile conditions, after opening the lid of the sterile rearing container described in the first embodiment, eggs and / or first-instar moths are inoculated into the container within a short period of time. The eggs to be inoculated are not limited, but eggs that have undergone the sterilization step described above are preferably inoculated. Furthermore, the first-instar moths to be inoculated are not limited, but preferably are within three days, two days, or one day of hatching at temperatures between 20 and 25°C, and within seven days of hatching at temperatures between 10 and 15°C. Furthermore, first-instar moths that have hatched but have not yet nested and / or fed are preferably inoculated. The number of eggs and first-instar moths to be inoculated is an appropriate number based on the capacity of the sterile rearing container, the amount of nesting material and artificial feed placed within it, and other factors. Therefore, if the eggs obtained after the sterilization step are in the form of an egg mass, they should be pre-adjusted to a predetermined number before inoculation.

[0169] The inoculation site is not limited, but is preferably on the nest material or near the contact boundary between the nest material and the artificial feed. This is because the hatched moths first start to build nests, so the burden of movement of the moths is avoided or reduced.

[0170] After this step, the opening of the sterile breeding container is securely sealed with a lid to maintain the sterile condition. The breeding container is then removed from the sterile conditions and placed in an environment suitable for the growth and development of the moth (e.g., 23°C to 27°C, which is the optimal growth temperature), where it can be left as is.

[0171] In principle, the rearing period in the sterile rearing container is a period pre-set in the rearing container based on the capacity of the heat-resistant container and the amount of nesting material and artificial feed. If it is the basic structure of the sterile rearing container for the moth described in the first scheme, it is constructed in a way that it can be reared from the first instar just hatched to the fourth instar without the need for feeding management such as bait replacement. Therefore, roughly this period becomes the rearing period in the above-mentioned rearing container. This rearing period is usually, for example, 5 months, 4.5 months, 4 months, 3.5 months, 3 months, 2.5 months, 2 months, 1.5 months or 1 month. However, depending on the remaining amount and / or state of the nesting material and artificial feed in the container, the amount of feces inside the container, and the density of the moth, the rearing period can be shortened or extended appropriately taking these into consideration.

[0172] In principle, nesting material and / or artificial feed should not be added after the start of breeding. However, they can be added to the container as needed. In this case, after sterilizing the outer wall of the sterile breeding container with 70% ethanol or the like, nesting material or artificial feed, which has been sterilized separately under aseptic conditions such as in a clean bench, can be inserted through the container opening.

[0173] On the other hand, after the start of rearing, the moths within the sterile rearing container are not, in principle, removed until the rearing is complete. However, some individuals may be removed as needed. In this case, after the outer walls of the sterile rearing container are sterilized with 70% ethanol or the like, some moths are removed under aseptic conditions such as a clean bench.

[0174] In addition, it is generally not allowed to put the moth worms once removed from the container back into the sterile breeding container.

[0175] 4-3. Effect

[0176] The aseptic rearing method of the present invention allows for the rearing of large numbers of 100 or more moths from the time they hatch to the fourth instar, without requiring any care, such as changing bait. This significantly reduces the labor, working time, and rearing costs associated with mass rearing of moths.

[0177] Furthermore, according to the aseptic rearing method of the present invention, the moths are reared under aseptic conditions, and therefore the risk of infections caused by bacteria, filamentous fungi, etc. can be infinitely reduced even in large-scale rearing.

[0178] Furthermore, according to the aseptic rearing method of the present invention, early stage moths that are difficult to handle can be reared without applying external physical stress such as contact, thereby reducing the mortality rate of early stage moths.

[0179] Example

[0180] <Example 1: Production of a Sterile Rearing Container for Corydoras Moths>

[0181] (Purpose)

[0182] The sterile breeding container for the moth of the present invention is manufactured.

[0183] (method)

[0184] Here, a method for producing a sterile breeding container capable of breeding 100 to 150 cricket moths is exemplified.

[0185] A 350 mL Tupperware (registered trademark) was used as a heat-resistant container, a 2 mm thick cork sheet was used as the nest material for the moth, and Insecta (Japan Agricultural Industry Co., Ltd.) was used as the artificial feed. Insecta paste was applied to a portion of the cork sheet. The cork sheet was placed at the bottom of the heat-resistant container. After the Tupperware lid was placed, the container was sterilized by autoclaving at 121°C for 20 minutes. The heat-resistant container with the nest material and the like after sterilization was used as the sterile breeding container for the moth of the present invention.

[0186] <Example 2: Aseptic rearing from eggs of the moth>

[0187] (Purpose)

[0188] The aseptic rearing of sedge moth eggs using the aseptic rearing container for sedge moths of the present invention was verified.

[0189] (method)

[0190] The moth used was the large-nested moth. Egg collection was performed using the method described in Japanese Patent Application Laid-Open No. 2022-092616. The eggs were kept at 24 ± 1°C and, two to three days before hatching, were surface sterilized by immersing them in 70% ethanol for 30 seconds. The eggs were then washed twice with sterile water. Eggs that were not sterilized with ethanol but washed twice with sterile water were used as a control.

[0191] The inoculation of eggs was carried out in a clean bench. The above-mentioned eggs were inoculated at a rate of 100 to 130 eggs per container on the nesting material in the sterile breeding container prepared in Example 1. Then, the lid was closed and placed at 24±1°C without changing the bait to maintain the sterility of the container. In addition, 59 containers inoculated with sterilized eggs and 90 containers inoculated with non-sterilized control eggs were prepared. After the sterile breeding containers were placed for 30 to 105 days, the nesting status, the contamination rate based on the production of mold (filamentous fungi) in the container after a specified period, the survival rate of the moth, and the relationship between the breeding period and the age of the larvae were verified. The age of the larvae was determined by the head width of the moths taken out of the nest. Taking into account the sterility of the container and the impact of the stress on the moths caused by the determination of the larval age, each experimental group was carried out in its own different container.

[0192] (result) Nesting state The moths that hatched 2 or 3 days after inoculation immediately scraped off the surface of the cork and began to build their nests. Figure 1 As shown, the moth forms a cone-shaped nest wrapped with cork particles.

[0193] Contamination rate

[0194] Thirty days after egg inoculation, the number of containers with mold growth anywhere inside was counted, and the contamination rate was calculated. The results showed that the contamination rate for the containers inoculated with unsterilized control eggs was 43.3%, while the contamination rate for the sterilized eggs was only 1.7%. This result demonstrates that surface sterilization of inoculated eggs is effective in preventing contaminants from entering the culture medium during aseptic rearing of sedge moths.

[0195] Survival rate

[0196] The average number of surviving moths within the sterilized egg inoculation container 33 to 35 days after egg inoculation was 119. Since the average number of eggs inoculated per container was 120, this suggests that the survival rate of early-stage moths when reared using the sterile rearing container for moths of the present invention is nearly 100%. It is generally known that moths have low adaptability and resistance to the environment shortly after hatching, and therefore have a low survival rate even in their natural state. For example, even in a laboratory under suitable rearing conditions, with fresh evergreen branches and leaves, the average survival rate of early-stage moths was only 30.2% 30 days after hatching, further decreasing to 29.4% at 33 days. Therefore, it is clear that the sterile rearing method using the sterile rearing container for moths of the present invention is extremely effective for efficiently rearing early-stage moths.

[0197] Relationship between rearing period and larval age

[0198] The age composition of the moth in each container 45 days, 90 days, and 105 days after egg inoculation is shown in FIG. Figure 2 After 45 days, the population consisted solely of 2nd and 3rd instar individuals. After 90 days, 4th instar larvae accounted for approximately 30%, with some 5th instar individuals also appearing. After 105 days, the proportion of 4th instar individuals increased to 45%, but the proportion of 5th instar individuals did not increase. These results clearly demonstrate that the aseptic rearing method using the sterile rearing container for moth moths of the present invention allows for the successful rearing of moth moths from the first to the fourth instar without bait replacement for at least 105 days.

[0199] <Example 3: Configuration of nest materials and artificial feed and survival rate of moth moths>

[0200] (Purpose)

[0201] The effects of different nest materials and artificial diets in sterile rearing containers on the survival rate and growth of moths were verified.

[0202] (method)

[0203] The basic operation in the manufacture of the sterile breeding container for the moth was the same as in Example 1. However, the nest materials and artificial feed in the sterile breeding container used in this example were as shown in Tables 1 and 2. Figure 3 The configuration of the four test areas shown in the figure. A box-shaped polypropylene container with a diameter of 10 cm and a height of 4 cm was used as the heat-resistant container.

[0204]

[0205] Ten newly hatched larvae were inoculated onto the respective nesting materials in containers in each experimental plot and reared at 24±1°C. The survival rate of the moths during this period and the average length of the long axis of the nests 15 days after egg inoculation were calculated. Since nest size is proportional to the body length of the moths, the measurement of the long axis of the nests can be used to estimate the growth rate of the moths.

[0206] (result)

[0207] Figure 4 The survival rate of the moth in each test area 15 days after inoculation is shown in the figure. Figure 5 The middle shows the average length of the long axis of the nest of the moth in each test plot 15 days after inoculation.

[0208] according to Figure 4In test plots C and D, where the nest material was kept in contact with the artificial diet, the survival rates after 15 days were 80% and 70%, respectively. In contrast, in test plots A and B, where the nest material was kept separate from the artificial diet, the survival rate after 15 days was only 20%. These results suggest that placing the nest material in contact with the artificial diet in sterile rearing containers can prevent the death of first-instar moths after hatching and maintain a high survival rate.

[0209] In addition, according to Figure 5 Fifteen days after inoculation, the length of the long axis of the nests of the moth was 5.34±0.41mm (n=7) in test area D and 3.64±0.33mm (n=8) in test area C. The surprising result was that placing the nest material in contact with the artificial diet also accelerated growth.

[0210] The above results suggest that regarding the configuration and positional relationship of the nest material and artificial feed in the sterile breeding container for the moth, the configuration in which the nest material is placed on top of the artificial feed and they are placed in the container in a state of contact with each other is the most preferred in terms of the survival rate and growth rate of the moth.

[0211] <Example 4: Aseptic rearing of first-instar moths>

[0212] (Purpose)

[0213] The aseptic rearing of first-instar bagworms using the aseptic rearing container for bagworms of the present invention was verified.

[0214] (method)

[0215] Larvae of the large-nested moth were used. Eggs were collected using the method described in Japanese Patent Application Laid-Open No. 2022-092616. The resulting eggs were incubated at 24±1°C. Within three days of hatching, scales and egg shells from the mother moth were removed from the body surface of first-instar moths that were not nesting or feeding at this temperature using a sterilized pen.

[0216] The inoculation of the first-instar moth worms was carried out in a clean bench. The first-instar moth worms were inoculated at 100 individuals / container on the nest material in the sterile breeding container prepared in Example 1. Then, the lid was closed and placed at 24±1°C. The bait was not replaced and the container was placed directly while maintaining the sterility. In addition, 27 containers inoculated with the first-instar moth worms were made. The artificial feed in the sterile breeding container was consumed by the moth worms in the container. When it was visually confirmed that it was almost gone (after 73 to 90 days, an average of 79.8 days), the container was opened, and the nesting state, the contamination rate based on the production of mold (filamentous fungi) in the container after breeding, and the age composition of the moth worms after the breeding period in the container were verified. The larval age was determined by the head width of the moth worms taken out of the nest.

[0217] (result) Nesting state Immediately after inoculation, the first-instar moths scraped away the surface of the cork and began building nests. Similar to the case of egg inoculation, the moths formed conical nests entwined with cork particles.

[0218] Contamination rate

[0219] Approximately 80 days after inoculation with first-instar moths, until the containers were opened, no containers developed mold inside. This result clearly demonstrates that if the moths inoculated in the sterile rearing container for moths of the present invention are at least first-instar moths, the sterile state within the container can be maintained until the bait in the container is consumed and depleted.

[0220] Age composition of the moth after rearing in containers

[0221] Approximately 80 days after inoculation with first-instar moths and opening the containers, the age composition of moths in each container was characterized by the largest number of 3rd-instar (46.8%) and 4th-instar (42.6%) individuals, with some 2nd-instar (5.8%) and 5th-instar (4.8%) individuals also observed. These results demonstrate that, even when inoculated with first-instar moths, the sterile rearing container for moths of the present invention allows for sufficient rearing of moths until the bait in the container is depleted, even when inoculated with first-instar moths.

[0222] All publications, patents, and patent applications cited in this specification are hereby incorporated by reference into this specification as they are.

Claims

1. A sterile breeding container for moths, comprising Heat-resistant containers made of raw materials with a heat-resistant temperature of 100°C or more, Artificial feed, and Moth nest material containing bark, The heat-resistant container is in a sterile state. In the heat-resistant container, the artificial feed and the nest material of the moth are arranged in contact with each other, or are arranged at a distance such that moths can directly cross the nest material and the artificial feed.

2. The sterile breeding container according to claim 1, wherein the artificial feed is attached to a portion of the surface of the nest material of the moth.

3. The sterile breeding container according to claim 1 or 2, wherein the heat-resistant container has ventilation holes.

4. The sterile breeding container according to claim 1, wherein the bark is cork.

5. The sterile breeding container according to claim 1, which is used for moths from the first instar to the fourth instar. The sterile breeding container according to claim 1 , wherein the nest material of the moth is arranged at a position higher than the artificial feed.

7. A baited nest material for moths, wherein artificial feed is attached to a portion of the nest material comprising bark. The baited nest material for moths according to claim 7 , wherein the bark is cork.

9. The baited nest material for the moth according to claim 7 or 8, which is used for moths from the first instar to the fourth instar.

10. The baited nest material for moths according to claim 7, which is sterilized and packaged.

11. A method for manufacturing a sterile breeding container for a sedge moth, the method comprising: A step of placing artificial feed and moth nest materials in a heat-resistant container made of a raw material having a heat-resistant temperature of 100° C. or higher; a contacting step of bringing the artificial feed into contact with the nest material of the moth in the heat-resistant container; and After the interior of the heat-resistant container is sealed, a sterilization step of sterilizing the container is performed. 12 . The production method according to claim 11 , wherein the contact is a state in which the artificial feed for the moth is attached to a portion of the surface of the moth nest material.

13. The production method according to claim 11 or 12, wherein the bark is cork.

14. The manufacturing method according to claim 11, wherein the nest material of the moth is arranged at a position higher than the artificial feed.

15. A sterile breeding method for moth moths, comprising: An inoculation step of placing eggs and / or early-instar moths of the moth in the sterile breeding container according to claim 1 under sterile conditions. 16 . The sterile breeding method according to claim 15 , comprising a sterilization step of sterilizing the surface of the eggs of the moth before the inoculation step. 17 . The sterile breeding method according to claim 16 , wherein the sterilization treatment is a contact treatment with 70% ethanol or 2-3% formalin.

18. The aseptic breeding method according to any one of claims 15 to 17, wherein the first-instar moth worms are moth worms within 3 days after hatching.

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