JPH07503484A - Biological control of termites - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Biological control of termites Background of the invention The present invention relates generally to the field of biological control of harmful insects, and specifically to the control of termites. This invention is in the area of using behavioral modifiers in combination with pathogenic fungi.

Termites cause damage to wooden structures, including homes, commercial buildings, and furniture. It also causes large economic losses due to the deposition of soil and mud during its invasion process. . Termites also attack carpets and other floor coverings, wall coverings, stucco, siding, and Damage to wall materials. Termite infestations in homes can be devastating for homeowners, especially those who sell their homes. This is extremely troublesome for those who are trying to avoid it.

The most common means of termite control are regular chemical insecticide sprays or injection of large amounts of pesticides into the soil surrounding buildings. The problem with chemical pesticides is that its long-term effects on humans, pets, and the environment are unknown. and; as chemical treatments are repeated, resistant insect species tend to emerge, making it difficult to kill insects. the need to further increase the amount of agent or use a different chemical; The chemicals currently used repel termites rather than kill them. It turned out to be less effective in termite control than expected.

Insect pathogens become more virulent and commonly used for the control of harmful insects. Can be used as a substitute for scientific insecticides. Fungi as biological agents for termite control Examples of potential insect pathogens suitable for use in

Fungi are found either as unicellular organisms or in multicellular colonies. fungi are true are nuclear organisms and are therefore more highly differentiated than bacteria, but how advanced plants are Not differentiated.

Fungi cannot use light as an energy source and are therefore It is limited to its existence as a living organism. The most common forms of fungal growth and reproduction is vegetative growth or asexual reproduction, and this reproduction involves sporulation and then germination of the spores. including. Asexual spores or conidia are formed at the tips along the edges of the hyphae and are multicellular. Branches into fibrous structures of the colony. In a favorable environment, conidia germinate and grow large. and form a germ tube. The germ tube develops into a hyphae over time and then into a colony. Form the knee.

To date, a large body of research has evaluated pathogenic fungi for biological control of harmful insects. Many are focused on applications involving harmful insects and mosquitoes that are important in agriculture. Meta rhizium aniso 1iae is the most widely used for biological control of insects. This is an example that has been extensively studied. M, aniso 1iae, by many methods These methods include direct spraying, inoculation, and injecting into harmful insects. refers to the application of fungi to plant or soil material on which insects live or are nourished. include. Fungal infections have been shown to be fatal in certain insect species. Fungi M, 　　lie is HanelanISOa and Watson, 73 Bull, Ent, Res, 305 -313 (1983) and Beauveria ba ssiana is 16 Entomo ha by Bao and Yendol Infect termites, as reported in A. 343-352 (1971). This is an example of fungi. In certain species of termites, Kraa+m et al., 40 J, Inv. As reported by ert, Pathol, l-6 (19B2), fungi can infect infected individual termites transmit the fungus to uninfected termites in their colony. Ru.

A limitation of most previous studies using fungal pathogens of insects is that insects are The research was conducted under laboratory conditions, which were quite different from those used in the study. Big In some reported cases, insecticidal treatment of treated insects may be due to ingestion of large amounts of spores or is achieved by inoculation, and the spores can be toxic if they enter the body, and Can be toxic by itself. In other cases, test tubes containing large amounts of fungal spores or or by rolling the insects or sprinkling them with fungi in a bedding dish. I infected it. Commercial or large-scale use of such methods is Obviously impractical. In addition, government regulations limit insect invasion areas, especially humans and food sources. Fungal pesticides that require random mass release of fungal spores where things can become contaminated can make registration difficult. Management and biological management of termites that invade homes and buildings For reliable and commercially viable control of termites, Methods to ensure a widely distributed fungal inoculum in a growing population are still being developed. Not yet.

Gunner et al. U.S. Patent Nos. 5,057,315 and 5.057.316 issue discloses the use of infection chambers containing fungal pathogens and attracting insects with fungi. The insects are then infected and killed. Examples include cockroaches and flying insects. 1nsect). The infection chamber design maintains fungal cultures and insects come into contact with this fungus.

Depending on the insect species, the chamber morphology, food, attractants, fragrances, aromatics (sce Attract insects to the chamber by one or more means, such as nts) or pigments. do.

It is desirable to provide an infection chamber for termites.

However, such chambers have never been previously disclosed, and termites vs. what form of such a chamber or what is used as an attractant It is unclear whether this can be done.

Therefore, it is an object of the present invention to biologically control termites using pathogenic fungi. An object of the present invention is to provide a method and apparatus.

Another object of the present invention is to provide a simple and reliable method for biological control of termites. and to provide an economically viable device for applying fungi.

Another object of the present invention is to prevent termite invasion and termite invasion by repelling termites. It is an object of the present invention to provide methods and means for preventing damage to structures or areas.

Another object of the present invention is to eventually eliminate all or most of the white loa in the breeding colony. The purpose of the present invention is to provide methods and means for infecting viruses.

Still another object of the present invention is to infect or kill termites with various fungi. The aim is to provide methods and means for .

Summary of the invention A method has been developed to protect structures from termite attack.

In a preferred embodiment, the termite is infected with the fungus GloeoflutrabeU or its volatile products, such as feeding, arresting, trailing A convocation prompt that triggers a series of actions, including trail following. Using a gift (herein referred to as an "attractant"), M, aniso 1i Fungi, which are natural termite repellents, such as A. ae., are used, and the Used to prevent termites from invading areas covered by Solitary or attracting fungi pathogenic fungi in combination with infection chambers, or prepared with nutritional compositions Provided in formulated form and placed in habitat corridors or corridors or termite nests Administered directly to initiate infection, transmission and colony killing. The benefits of fungi are Once established, the fungus continues to protect structures from termites indefinitely. That's true. The two most preferred pathogenic fungi are Metarhtzium and iso l1ae and Beauveria bassiana, but white When the ants are inoculated through the infection chamber, they also use other fungi that are pathogenic. obtain. L'' Wind l■■ is a suitable repellent fungus. Gloeo h Ilu+g trabeuIll is a preferred attracting fungus. Examples include M, aniso of termites using a chamber containing B. 1iae and B. bassiana. Show control.

Brief description of the drawing FIG. 1 is an overall view of a termite infection chamber.

Figures 2A and 2B are graphs tracking the decline in termites after exposure to fungi. The vertical axis shows the survival rate in percent, and the horizontal axis shows the elapsed time after treatment in days. vinegar. Figure 2A shows survival after exposure to L'' decoy 11■ compared to control (circle). Figure 2B shows the control (diamond); The survival rate (squares) after exposure to ana is shown.

Detailed description of the invention The present invention is essentially based on two discoveries: (1) that pathogenic fungi , which can be used to infect and kill termites, but does not allow termites to come into contact with the fungus. (2) an attractant is required to induce the particular type of bacteria to Due to its inherent repellency to termites, It can be used to repel.

Pathogenic fungi may be used alone or with attracting fungi or compounds derived therefrom. , or in combination with repellent fungi, to the area to be treated or protected. agents, or fungi for termite infection or repellency that may be administered in an infection chamber. Start culturing the species.

The main problem with pathogenic fungi for termite control is that pathogenic fungi Termites are repelled, resulting in a well-spread infection and a significant reduction in termite numbers. It is extremely difficult to reduce the amount. This problem is caused by the avoidance of pathogenic fungi. Combining fungal or other attractants with pathogenic fungi to overcome the It was overcome with.

The other major issue is how to maintain fungal cultures that effectively kill termites. child The problem is that when the formulation is administered to soil containing termites, it is difficult to establish a culture of the fungus. This was overcome by developing effective nutritional formulations. The above nutritional preparations are On a nutrient medium such as bran or as a dry powder such as grain or sawdust It consists of fungi on a cellulose-based food that can be administered.

Yet another major problem was how to transmit the fungal lethal infection. This problem , an effective infection to establish a fungal culture in the vicinity of the area to be treated or protected. This was achieved through the use of a chamber. The infection chamber contains the nutrient source for the culture and contain the bacteria before they have established themselves on the site to be protected or treated. and protect it.

Bacteria selection Useful fungi are found on infected termites (pathogenic fungi) or termite-infested trees. as isolates from wood (attractive fungi) or from soil using conventional techniques. can be obtained as an isolated bacterium or from preserved cultured collecticons. this These bacteria are American Type Culture Co11ectio. n, 17.301 Parklawn Drive, Rockville Le, Maryland (ATCC), or USDA-ARS medium for pathogenic fungi. Nutrient collection, U, S, Plant, 5 oil andllater Lab, Tower Road, [thaca, New York (A RSEF), etc., and these can be used without restriction. Soil isolates are , screen soil samples for the presence of fungi and then test for insects in the garden. This can be obtained by examining the effect using a bioassay. Pathogenicity exposed to fungi Proven at the time the insect dies.

There are two types of pathogenic fungi, M. basiana and L. bassiana. , was shown to infect and kill termites.

Examples include MM, anis Dan11 [ae], which was also shown to repel termites. Contains ATCC62176 and ARSEF 1112.

Useful fungi can be obtained by incubating termites under laboratory conditions as described in the Examples below. can be obtained by screening soil isolates by exposing them. Yes The most common pathogenic fungi are those that kill termites after direct contact. useful death Evasive fungi inhibit termites in soil box tests as shown below. It is a fungus that is repelled. As used herein, the term "repellent fungi" refers to % etarh Oisan 1 Yuwang, or the compound produced by this fungus that effectively repels termites Contain things.

TojjL 珂 Useful attractants can attract termites, irritate termites, or otherwise attract termites. It is an attractant that attracts members of the population to congregate and feed. Termite A more preferred attractant is Gloeohllum trabeum (formerly known as Lenzites trabea) and certain of its metabolic products, extractable Wood decayed by specific fungi containing attractants and the following traces: il) Contains specific organic compounds that mimic pheromones: Watanabe and Ca5ida, 56:3 J, Econ, Ent, 300-30 7 (1963), Matsumura, F., et al., 65:2. L, Eco n, Ent, 600-602 (1972), 5aythe, R, V, et al. 58:3 J, Econ, Ent, 420-423 (1965), Nara Prestwich, G. D., et al., 10:8 Journal of Cheo+1cal Ecol.

■1201-1217 (1984). These teachings are incorporated herein by reference. used. Chamber shape and chamber composition, and chamber location can be used for termite attraction.

Wood that has been decayed by specific fungi is treated by Matsumura F, et al., 62:3. J, Econ, Ent, 699-603 (1969); Watanab Attractants for termites were added as described in E and Ca5ida (1963). contains.

For example, wood decayed by the brown rot fungus G, trabeum (S, S , E)-3,6,8-)” ``Tekatorien-1-ol and termites Contains other compounds known to do so. Grace, J., 92 Proc, Entomo! , Soc, Wash, 773-777 (1 990) was decayed by G. trabeum in combination with antioxidants. Includes termite trail following activity in response to extracts from wood , performed behavioral bioassays. Watanabe and Ca5ida (19 (63) also found that many organic compounds can attract termites. Shiro Ant-attracting activity is influenced by wood size, and size guarantees termite recruitment It was observed that the minimum mass must be exceeded.

At least one species of mG, trabeum, attracts termites and is pathogenic. It has been shown to produce substances that mask the repellency of fungi. Other should be useful Fungi are fungi that cause wood to rot. Some isolates of G. trabeum Available from ATCC; ATCC 13021 has been tested and is attractive It was confirmed.

Rin 9” U1 Kamei Suitable culture media that can be used within the chamber are known. known to those skilled in the art , and examples of commercially available culture media include % Metarhizium and Potato dextrose agar (PDA) or Co. Contains rice agar. Rice agar medium is 1% dextrose. loin, 1x yeast extract, 5% rice flour, 1.5% agar, and 0.5% 5x Consists of Dubois spore-forming salts. 5x Dubois sporulating salts, 1 5g (NU-hsOa/10100O; 0.30g Mg5Oa 7820 /1000! ! I; 0.15g Mn5OaH20/1000m1; 0 ．． 0375g CLISo, 5H20/1000111; 0.0375g Zn5Oa 7H2/1000+1+1; and 0.0038g Fe50 47H20/1000111. In winter, ensure that the salt is completely dissolved before adding the next salt. The solution is then autoclaved.

Other useful culture media are known and may be optimized from culture media known to those skilled in the art. It can be done. For example, either PDA or a simple cellulosic material like sawdust can be used as a culture medium for G, trabeu+m growth.

i Imperial EZ Bani As used herein, the term "chamber" refers to the initiation of an infection process within a population. refers to the inoculation chamber used to The infection chamber is (1) emergency condenses a very high concentration of pathogenic fungal inoculum into a small space and infects termites. and bring the termites into contact with the spores to be killed; (2) Eleventh spores and termites; / or contain mycelium, minimizing exposure of the fungus to the surrounding environment and keeping the fungus in a protect the fungal culture from contamination, thereby increasing the viability of the culture and preventing contamination of the fungal culture. and (3) sawdust and wood that serve alone or as a food source for termites. Gloeohl combined with a chip to overcome the repellent properties of pathogenic fungi Presence of termite attractants such as lumtrabeun+ and its volatile products This attracts termites to pathogenic fungi.

The chamber may be made of non-biodegradable or biodegradable materials and It is designed to establish an effective infection site for the infection of lizards.

Small, lightweight infection chambers (or “pegs”) are unobtrusive and can be easily placed in locations of severe termite infestation, increasing the effectiveness of the device.

A specific fungal culture may be used as an attractant within the chamber, and a second fungal culture may be used as an attractant. Similar cultures provide a continuous supply of infectious spores over an extended period of time. these The spores attach to the termite and produce germ tubes that penetrate the termite epidermis and remain for a short time. will lead to death. Attractants, such as volatile compounds produced by wood decomposition, alone or with wood-degrading fungi such as Gloeo h llum trabeum It may be provided in combination with The main means of infection is external contact, but Shiloa li can also be infected by contact with each other and by ingestion of spores.

These attractants can be used alone or in cellulose-based materials that serve as a food source for termites. In combination with the material, new termites between the chamber and the colony are Causes a behavioral reaction in the tracks, thereby making it possible for other termites to become infected It gathers its body and transfers spores to other individuals within the colony.

Once the termites enter the chamber, they come into direct contact with the pathogenic fungi. of this fungus The conidia attach directly to the termite's body. After attachment, conidia germinate on the insect epidermis. , forming a germ tube that penetrates the termite's integument. This germ tube is inside the termite. It continues its penetration into the body cavity (hemocoel) and kills the termite. vinegar. After the termite dies, given the right conditions of relative humidity and temperature, the fungus mycelium can sporulate on the termite's body, and other termites can be infected by exposure to conidia produced on the cells. Non-infected Shiroa When termites are exposed to spores on infected termites or termite carcasses, they become infected with pathogens. effectively transmitted. Transmission of spores is caused by termite social behavior such as: Reinforced: For example, blooming, communication through contact transport and burial of carcasses, and exchange of dung material, and cannibalism. ism). There are also termites that ingest the spores and contribute to their death. can be destroyed or killed.

The chamber includes an outer housing that forms the basic shape of the chamber, and Consists of an inner core mixture that fills the housing unit. chamber housing is , glass, metal, extrudable or moldable plastic, or wood. may be constructed of conventional materials, preferably cardboard, cardboard, wood, compressed Resin - wood chips or particle board, expanded cellulose, papier mache material (p apier mache) or a biodegradable cellulose paste such as peat. A small opening 12 (diameter 2-5II11) large enough to pass through constructed with material , and a thin wall 14 that termites can easily chew through.

A preferred embodiment is a tubular structure. The shape ranges from circular in cross section to multi-shaped with multiple tips. Although they can range up to square shapes, triangular tubular structures are more preferred. Chamber The length of the chamber ranges from 10cm to 30cs, and the width of the chamber is 3cm. The range is 8 cm from The chamber tip 16 accommodates placement of the chamber on the ground. The tip is pointed for ease of use. The chamber tip 16 is flush with the housing wall 14. Can be made of one material and coated with resin or other materials to increase strength or the tip may be made of a harder material such as plastic or wood. Ru. The top 18 of the chamber housing is covered with a paper or plastic cap after filling. covered with a cup.

Inside the housing, the core of the chamber is filled with a mixture of several components: Obtain: (1) Basic food source for termites (cellulose paste material, cheapest (2) Attractant: 4-8 week old G,trabe grown on the sawdust layer culture of um, G, killed culture of trabeum, or G, trab Extracts of eu+i cultures or synthesized chemical attractants and restraints (arre stan (formulated wet or dry with food source and protective agent f=M, anisoli or other pathogenic organisms that grow on solid or liquid media. microorganisms, or spores). The core mixture was inoculated with trabeu+a. 100g of scraps: mixed with oat bran and formulated with dried mycelia A suitable ratio of 5 g: M, atop y1 ■ 1 g of spores, with a proportionate excess of lethal fungi. It should contain an attractant.

These components can be spatially separated or homogeneously mixed and are Maximize contact with deadly fungi and reduce overload of deadly fungal spores to individual termites. Provide a surplus dose. The culture may be applied to an inert grid or grid, where , the lattice or grid is suitable for growing or placing fungal cultures. , or alternatively, a supporting matrix for retaining nutrients within the device.

The chamber may be properly installed as follows: (1) the chamber is cured; (2) when constructed of materials, driven directly into the ground; (2) excavated with a tool such as a trowel; (3) push the chamber into the hole or core sampling device; Place in the inner sleeve of a hollow tool that can be used to insert it into the ground.

The following are the topography, termite density and type, climate, and chamber effectiveness. This is a general guideline that may be modified depending on other variables that may affect π. Chan The bar shall be 3.0 meters away from this structure at a perimeter around the area where the structure is to be protected. It is located at a distance of at least 5 to 10 meters away, preferably 5 to 10 meters away. chamber – are the areas where termites are active or where termites are active and the areas to be protected. should be installed centrally between the The chamber is 0.1 meters to 1 They should be placed apart, with a distance of meters.

Greenhouse and laboratory tests have shown that the lethal bacteria M, aniso 1iae are present for over 90 days. B. beauvern remains in the soil at high concentrations for a long time. It also shows that it remains in agricultural soil from May to October.

, a dyeing chamber and a Fukuin product. An infection chamber suitable for infecting termites is initially Build the housing unit in and then fill the housing unit with the core contents It can be constructed by The housing unit is made of flexible, flat cellulose It can be cut from the product sheet, folded into the desired shape, fitted and glued together. Ru. First, plastic, resin/wood, papier-mâché material before cutting and folding. It is more desirable to mold it from peat or peat. Molding is wall and foldable Grooves and holes are formed to create the final shape. Molding a complete shape at once, Rui can be molded into sections that can be assembled with glue, with holes drilled. Make a dent.

The assembled housing is then filled with core mixture. The core mixture is 3 It is formulated in stages. In stage I, cellulose-based nutrients, typically sawdust, are added. Debris is prepared for termites and G. trabeum. Sawdust is Place in a plastic bag or other container and allow a 12-24 hour cooling period between each sterilization. It is then sterilized twice in an autoclave. Potato dextrose meat on sawdust Inoculate with G. trabeum mycelium recovered from a liquid culture medium such as juice. . Mix sawdust and G. trabeum mycelium, and at 24-27 °C for 4. Culture for 8 weeks.

In step ■, the cellulose G, trabeum culture is mixed with the formulated M, an Mix with iso l1ae hyphae. This formulation was prepared using a liquid medium (1% yeast extract, 1% % peptone, 4% dextrose), aniso l1ae The mycelium is mixed with a dry food source (oat bran flour), mineral salts and protectants. It is prepared by

The mixture is dried and ground to form granules.

Finally, in step (2), the above preparation is mixed with L''' top 1 spores. The above y superparaconidia, from rice agar growth plates after 1 to 3 weeks of growth, or , collected from the bag of inoculated rice particles. Then the housing , filled by hand or machine with the core mixture and sealed with a cap. a Alternatively, a support matrix core or grid core may be inserted within the housing. Ru. The matrix core has pockets filled with the fungal preparation as described above. Here, the above fungal preparation refers to G on cellulose paste, trabe■ and M, antμ technology on agar, or dried mycelial preparations or These are the collected spores.

The following is an explanation of the efficacy of infection chambers, formulations and their components in termite control. Figure 2 is a non-limiting example showing theoretical and functional parameters. in all cases It has been observed that termite behavior is significantly influenced by chamber components. and the termite population is greatly affected by the fungi present in the infection chamber. Diminished.

Example 1: Direct exposure to Metarhiziuo+ and Beauveria Weed Wood Termites (Zooter+ao sis an us ticollis) demonstrates the efficacy of the fungus in killing termites.

Termites in groups of 5 were transferred to 7 day old growth fungus culture plates.

After 5 minutes, termites were transferred to a clean plate and excess conidia were removed.

After 30 minutes, transfer the termites to a holding plate with a layer of moist filter paper and Kimwibe. did. Then, place the holding plate on a plastic plate with a damp paper towel at room temperature. I placed it in the box. The results are shown in Table 1.

(Margin below) Table 10 Termites (Zooters) after contact with selected fungal cultures is a usticollis) Observation of exposure for 30 minutes After 3 days After 5 days Later termite carcasses 1 look at the appearance Kei Soguchi ATCC62176(s) is covered with 100% bacteria. low) B. ESC71 covered with bassiana disease 1aoxm (sick ) B. bassiana disease 80% (Rakano fungus ARSEF 721 ( sick) is good growth control 0 pieces Not applicable (well) (not applicable) After 3 days as listed in Table 1 , #62176 and #71 all died and the control All of the termites in the area survived. After 5 days, the carcasses were covered with fungal growth.

Direct contact of an individual termite with a pathogenic fungal culture results in the death of the termite. It can be concluded.

Example 2: Eastern 5ub by direct exposure of termites to fungal isolates The feeling of terranean termites (Reticular termes) Dye.

The termite-containing wood was removed from Montagu, Massachusetts. e) were collected from the forest land. Separation of termites from wood and bioassay Eight colonies were formed from which individual termites could be extracted. 10 termites, spores The vitriculture containing the pathogenic fungi forming was left on the surface for 15 minutes.

After treatment, termites are transferred to a holding plate containing filter paper and observed. did.

From the results shown in Table 2, it is clear that Matarhizium and Baeuveria That certain strains are highly effective against termites and can be used for termite control is clear. Soil isolates also showed high potency, indicating that these pathogens The present invention shows that isolates of sexual fungi can be obtained from soil using conventional methods.

Example 3: Colony infection by directly releasing infected termites into the colony infection of the chamber.

Four groups of termites (Z, an usticollis) were treated with cellulose powder. M, antm1 pair or B scraped from the agar growth plate. , a small piece of wood coated with a mixture of Bassiana conidia (2c+lx O, ScmxO, 5) was placed in a Petri dish. Infect termites for one hour source and transferred to a fixed plate. After 48 hours, the exposed termite One was introduced into a group of other termites that had not been exposed to the fungus. 9 days after contact A second group of termites, which showed no signs of disease, were left unexposed to the fungus. The termites were then transferred to a third group of termites. Over time, each of these groups of termites Observed death.

Five days after initial exposure to the fungus, members of the first group of termites in both dishes - also died out. The second group died on the day between 13 and 15. 14 days since last move After 23 days, all the exposed termites had died, and some had grown fungi. It was long.

All control termites were alive in each group.

The results of this study show that pathogenic fungi can be transferred between termites and the original source of infection. It was shown that this caused the death of termites that did not come into contact with it at all. Transfer of infection is probably This probably occurred during a social behavior such as blooming.

(Margin below) Table 2: Reticulitermis ■■n■ by contact bioassay 1 Fungal activity against Example 4: Infection of termites by exposure to fungi outside the nest.

3 small vetri dishes (15x100 m+g) and 1 large plate ( 20x150 + am) are connected with tubes to create a series of internally connected chambers. It was. Large plates with a small amount of wood to imitate water reservoirs and nests has. Termites were allowed to survive in this environment for one month. Conidia (L”jy and B or B. bassiana) by applying the fungal preparation to the filter. cover on the tar paper disk and/or toothpick placed in the outermost chamber. and applied it. Each unit had 6 termites and was treated twice.

The results are shown in Figure 2. Data from duplicates were combined to increase the number of samples . In the case treated with ATCC62176, LT5o died in 26 days; However, all controls survived.

Similar results were obtained when the experiment was conducted with ESC71, with LT5° of 22 days.

The conclusion from these studies is that the formulated fungi should be placed within the foraging range of termites. (and all members of a small colony of termites are killed by fungal infection) It means getting.

Example 5: Applied to the soil to infect termites at sites remote from the nest. Comparison of fungal preparations.

1. Soil box trial Fill the bottom of a plastic box (15x20x35 am) with 2-3 cm of soil. Filled. At one end of the box is a mock-up with 300 termites. A nest was placed. This nest was constructed from the bottom of two Petri dishes (20 x 100 ml). The side is cut to have four holes, placed with the surface open, and the fie Filled with router paper and pieces of wood. Termites can adapt to their environment within 3 to 10 days. It was. At the opposite end of the box, dig a hole in the soil 1.5-2 cm deep. , and applied fungal preparations. Wood block (5ClX3 ClX7 C1) was placed in the hole containing the formulation and half buried in soil. The formulation consists of (A) particle shape (o oat bran grains and mycelia) and (B) powder form (cellulose and It consists of conidia and conidia). For wood, soil, and nests, The activity of fungi was periodically tested.

The results shown in Table 3 indicate that wood can be protected by applying fungal preparations in its vicinity. showed that it can be done. In addition, termite colonies are located far away from the nest. that ants can be killed by applying a formulated fungus to the areas they attack. Indicated.

(Margin below) Table 3: Treatment of termites made of soil and heat-treated fungi ATCC62176 Formulation A: Fungal growth, all termites are killed. Formulation B: No growth, all termites are killed. ESC71 Formulation A: Fungal growth. There are no termites near the wood, and the nest is alive. Formulation B: No growth; termites are everywhere, and the nest is still alive Control Formulation A: Termites dig tunnels under the wood. , make the wood full of tunnels Formulation B Termites dig tunnels under wood everywhere. , make the wood full of tunnels Example 6: Testing of fungal pathogenicity induced remotely from termite colonies.

Fill the bottom of the plastic box (LSI 20!35 cm) with 2 to 3 cIl of soil. filled with. An artificial nest with approximately 300 termites was placed at one end. nest is constructed from the bottom of two Petri dishes (20xlOQ + am), with cutouts on the sides. It had four holes depending on the cut. Place the two bedding dishes with both sides open. and filled with filter paper and wood chips. Move the termites to the nest, and within 3 to 7 days adapted to the environment. At the other end of the box, a hole is dug in the soil and dried mycelium is prepared. Apply the agent and place a wood block (5x3x7 am) in the hole to loosen the soil. Filled the rear. For wood, soil, and nests, fungal activity against termites The sex was checked regularly.

The effects of a group of isolated bacteria are shown in Table 4. In most cases the fungi have grown well. In certain cases, termites were repelled from areas where the fungus grew. in a limited number of cases The termites were dead inside the nest. The fungi were then collected from the nest carcasses.

The most effective isolates for killing termite nests are Metarhi, zium and ATCC62176 and ESC70 with Met. Other highly effective and repellent isolates are 2080 and 111 It was 2.

(Margin below) 1, -'f:p2tono-7'f'n'l1LktItlip9,l2#15 1+:r(bfesne3r)7j4 Example 7 d Test of termite attractant.

For short t1 distances to determine the effectiveness of G, trabeui as an attractant. conducted initial research. Arena made of glass and wood (60x60x5) c+s) was covered with a thin layer of plaster. In the final experiment, add a thin layer of soil Place it on the surface of the tar. Has 6 holes on the side and has moist filter paper , place termites (150-300) in an upside-down dish on one side of the arena. Placed.

As one set of experiments, (a) a potential attractant and (b) a control material A centrifuge tube (1,5 ml) filled with liquid is separated by 15 am and then and placed 25 am away from the nest on the other side of the arena. arena were video recorded by a time-exposure camera and observed for 15 days. Another experiment A similar set-up was made for the second node of G, trabeu+n. - sawdust, and M, aniso l1ae (conidia or hyphae) and G , trabeum-sawdust mixture is G, trabeum-sawdust and Control was used in place of sawdust to fill the tube. all fruit The experiment was repeated three times.

The results show that sawdust infected with G. trabeum is more active than uninfected sawdust. Indicates preference. M, aniso 1iae conidia or mycelia G, t When mixed with rabeum, termites will react to the presence of M. anisolia. I don't respond. In other words, termites use the attractant tube and the attractant and Metarhiz. It invades both tubes containing fum. In both cases termites 2 to 7 hours after the test is set up, contact with the tube, and of sawdust, or G, trabeum/L' 1st wear-sawdust mixture. I started out. If soil is present, termites will tunnel through the soil towards the test sample. I dug it. M, aniso l1ae hyphae and rainbow trabeum-sawdust and G , trabeum - In a test comparing it with only sawdust, M, anis Nests exposed to o1iae will completely die after 15 days (100% mortality rate) . M, aniso l1ae and G, trabetrm - preparation of sawdust conidia did not achieve this high mortality rate.

The conclusions of these studies are that (1) termites are more susceptible to fungi than undecayed wood; It prefers wood that has been partially decayed by trabeum; (2) G. trabeus In the presence of G, termites ignore the deadly fungi and perform similar activities; This means that trabeu+i can suppress the repellent effect of lethal fungi.

Example 8: Establishment of termite foraging territory using attracting fungi.

Set up the arena (122x366x30.5 C+A) and covered with soil. 2 with about 1000 termites and filter paper The artificial nest in the bird dish was placed 61 cm away from three of the four walls of the arena. placed at one end.

A) G, a piece of wood (15x3x2 am) rotted by trabeun+ , 122.183 and 244 am away from the nest in the center of the arena. It was placed in These were left for one week, and termite activity was observed every other day.

B) G. A cardboard box filled with sawdust planted with trabeum and incubated for several weeks. Take the tube (height 15 crax diameter 4 c+g) from the nest to 122.183 and human. They were placed at a distance of 244 cm. Leave these for a week and check termite activity regularly. observed.

C) Move the above wood pieces and cardboard tubes within 60 c+a of the nest. Ta. These were left for one week and periodically inspected for termite activity.

Termites were observed to make holes 30 to 65 cm from the nest.

Individuals were observed within a range of 200011 from the nest. However, the tree in experiment A No termites were observed on, in or around the timber pieces or in the tubes of Experiment B. won. On the other hand, as shown in Experiment C, a piece of wood or a cardboard tube is placed towards the nest. When moved closer, termites can be observed overnight in wood chips and tubes. Ta.

The foraging area of termites may be more restricted in laboratory conditions than in the field. this Conditional muster only occurs when a limited distance is crossed. enough food in the nest Example 9: Attraction of termites to fungal attraction chambers.

A. Kokuhai Masae Approximately 1,000 termites were found in Montague, Massachusetts. It is obtained by separating it from wood collected in forest areas of placed in a vetri dish.

The test arena is 122X122X30c+a (length x width x height) and is covered It was made from plain plexi-glass. Arena is 5 Filled with cm of soil. High-pressure sterilized damp wood piece (15x3x2 c+a ), G. trabeum cultured for 8 weeks was planted, and the diameter was 4CI11 length. It was placed in a 13 cm paper tube. Similarly, a piece of high-pressure sterilized wood is G, tr The operation was carried out without planting abeum. In the third case, use a piece of wood that is not autoclaved. and filled the same kind of paper tube. termite colony covered with petri dish It was placed in the center of the arena. The four holes in the wall of the Petri dish are where termites can enter. Working in direction, each hole was oriented towards a corner of the arena.

The tubes were placed perpendicular to the arena wall, 3 on each side, and 10 c Each tube was partially inserted into the soil +a apart. The tube arrangement is as follows That's right.

Surface L: Tube A-G, trabeu++ wood tube B wood tube material Tube C-Wood Side 2: Chupu D - Autoclave sterilized wood Tube E-Wood Tube F-G, trabeum wood side 3: Tube Q-Q, trab eum wood tube wood tube water material Tube C-Wood Side 4: Tubego autoclaved wood Tube K-G, trabeui wood tube wood tube After the tube, the termites in the nest invaded the soil, but no termites were found in any of the tubes. No ants were observed. After 5 days, all teams with rainbow trabeun+ Termites were observed in the tube, but not in other tubes. .

The conclusion drawn from this experiment is that the G and trabeum in the chamber are Even when present, they are effective in attracting termites.

B. Sawdust test The experimental conditions for the wood test were the same as above except for the following description.

A variety of moist sawdust listed below was used in place of the wood pieces in the cardboard tube. . (1) Fresh sawdust (moistened), not autoclaved; (2) Autoclaved sawdust Trabeum sawdust (moistened and autoclaved for 25 minutes); (3) G, trabeum sawdust (Moisten, autoclave, and plant G. trabeus and culture at room temperature for about 8 weeks. The tubes used for the test were arranged as described above in the following order:

Side 1: To the tube - autoclaved sawdust Tube B - non-autoclaved sawdust tube (: -G, trabeu+m Saw layer side 2: Sawdust that has not been sterilized by tube-low pressure Tube E - sterilized by high-pressure scraps Tube F-G, trabeus saw layer side 3: Tube G-High pressure sterilized. No sawdust tube H - autoclave sterilized sawdust Tube I-G, trabeuve saw layer side 4: Chicove J-autoclaved saw waste Tube K-G, trabeum sawdust tube L-sawdust not autoclaved The results of this experiment showed that the termites in the colony were able to dig through the soil, leaving almost no residue in the nest. There was no such thing. Termites are all G's.

In the trabeum tube and one autoclaved tube (tube H) Observed. The conclusion of this experiment is that G. trabeum attracts termites, but , that only sawdust was not attracted in the time frame and distance used in this study. Met. Termites travel from a group of chambers placed in the soil. Only chambers inoculated with u- were selected.

Example 10: Comparison of fungal formulations in a soil box selection test.

Line the bottom of the plastic box with 3 cm of top soil (top 5 oil). did. An artificial nest with 300 termites was placed at one end. two nests Build from the bottom of a bedding dish (20 x 100 e■) and make 4 holes on the sides. , placed open-sided and filled with filter paper and wood chips. termites was adapted to the environment within 3 to 10 days. Place the two small blocks of wood in the box. It was placed in a shallow hole in the soil on the other side.

Each block is treated with an attractant (G, a piece of wood infected with trabeum) or a preparation. (ATCC 62176 mycolia mixed with oat bran) or its Treated with both or not treated.

The arrangement of blocks in each box is shown below.

(Margin below) Box Block 1 Block 2 ^　Wood　Wood and attractant B Wood Wood and formulated C wood and formulated 62176 wood and attractant D wood wood and attractant pulling agent and formulated 62176 Place the attractant on the wood block and add the formulated lethal fungi (62176) to the wood block. Buried in the soil below the block.

For box A, termites are more likely to have rainbow Lrabeun+ than wood-only termites. He preferred tomo wood. Termites are wood blocks placed on the wood block surface and preparation I preferred untreated wood blocks to wood blocks. These results are L'tan! Top ■ It also shows that it repels termites. In box C, termites are The wood block treated with G, trabeum was higher than the wood block placed on the standard preparation. I liked rock. M, antil has similar block (box D), the termite nest is placed in M, a like box B. It seemed to die out faster than the niso 1iae alone.

G, trabeum attracts termites to gather others, build a nest, and It has been shown that children can be fed with food. M, aniso liadan came into contact with termites. If you touch it, it will repel termites. Combining two types of fungi can kill termites. Contact with dead fungi was facilitated by the presence of attractants (recruitment stimulants). Therefore, G, trabeu+m, mixed with an appropriate ratio and spatial arrangement, is It was concluded that it suppresses evasiveness and eventually spreads the infection, leading to the death of the colony. Ru.

Example 11: Repellent effect of M, a supplementary ATCC 62176 granular formulation.

In the previously described arena, G, two Petris with trabeum sawdust Place the plates 45 cm apart from each other, then place them 60 cm apart. A termite colony was placed there. Width 20 mm made of culture 62176 preparation ring around one Petri dish and the other Petri dish as a control. Le 11J agent commercial, surrounded by oat bran (no oat bran).

Then mix the formulation with the soil and place the formulation area around the tray of trabeum sawdust. was formed. and spray water into the arena to set the soil and formulation. -I did. After 5 days, termite colonies were placed 60 cta away from both dishes. I moved it to Na. Growth of fungi and termites in the dishes of G. trabeum The activity was observed and used to evaluate the repellent effect of the formulation. Set up the same way as above. The 3 arenas that were popped are repeated experiments! , ■ and ■ set at the same time was pressed.

Twelve days after introducing the termite colony, the termites were transferred to the control formulation area. area, and also have G, trabeum in all three arenas. Reached the plate. There were several hundred termites in each of the three dishes. termites in blood He also dug through the sawdust and opened a tunnel that was visible from outside the plate. two at the same time Only in the arena (Arena! and ■), a small number of termites 62176 Passing through the formulation area, only 20 termites were observed on one side of the attractant dish.

Observe termite activity and tunnels from outside the Petri dish surrounded by 62176. It wasn't done.

Number of termites passing through the 62176 formulation area and control formulation area It is clear that there are significant differences between termite numbers. This preparation is G, t 62176 is effective against termites under the condition that it is spatially separated from rabeum sawdust. This shows that there is a deterrent effect.

Example 12: M, A as a granular formulation for protecting the wood structure of the arena Use of niso 1iae ATCC62176.

Gu Yanhang Keiyan Fangguang The arena is IZ2x366x30.5 cm (width x length x height) with a depth at the bottom. It was a plexiglass arena with 15+a+i soil.

Termites were collected in Montague, Massachusetts and kept at room temperature. Before use Separate the termites from the wood and transfer them to a petri dish filled with filter paper. Ta.

The timber frame was assembled using stock lumber.

Part A was prepared using four pieces of wood (5xlox30 Made with c+*). Nail these four pieces of wood together and make a 10x35x3 5 A rectangle of c+i wood was formed.

Part B was constructed from four pieces of wood (5X10X30 am). Nail these together A rectangle of wood measuring 5x40x40c+i was formed. Part B of Part A The head was nailed to form a timber frame.

ATCC culture 6217611 offspring collected from liquid culture were grown in sterilized oats. Mixed with bran flour. The mixture was dried overnight at room temperature with aeration, and then ground. Sieve through #14 sieve to form grains.

G, trabeum wood, wood pieces (13x3x2 c+a) are sterilized under high pressure, Ltrabeus and cultured for 4-8 weeks at room temperature.

A wooden frame was placed in the center of the arena. Culture AT around and under the base of the timber frame. The CC62176 formulation was mixed with the soil and a timber frame was set on top. in the center of the timber frame , G, trabeu+m A wooden pile was set in the soil. and , the wood frame was covered with a sheet of cardboard to prevent drying out and also to keep the arena dark. wood A termite colony is placed at each end of the arena 120 c+a away from the frame. (600-1000 individuals) were transferred together with a small amount of moist filter paper.

The colony is then covered with a Petri dish with a small number of holes on the side to form a termite nest. accomplished. One of the two termite colonies used had approximately 1000 termites. and the other had about 600 termites. and each arena for one week. Tested once or twice, wood with timber frame and G, trabeum The fungal growth and termite activity inside were investigated. It also adds moisture to the soil. I sprayed it with water regularly to keep it clean. Another arena made with control (oat formulation without 6217G mycelium), and 1000 seedlings each. A similar material was used, except that two termite colonies with termites were used. It was set up using materials.

Fungal growth was observed on the formulation two days after application. 1 week after application 62176 started to form conidia. After the start of production of conidia, production continues for about 20 days. The drug and conidia were observed. At the same time, the termites are in and around the nest. It was active in the soil, but also in the protected "wooden frame". It didn't even exist in the wood pile.

Five weeks after setting up the experiment, several hundred termites were placed under the “wood frame” and It was observed inside a trabeum wood pile. After 10 days, the wood frame It was observed that all the termites (several hundred) under “G, t The termites inside the "rabeu+a wood pile" were still alive.

2 after the termites move to G, trabeum wood pile” and “wood frame” After a week, the inside of the wood and the termite nest ( We observed dead termites in one of the original nests (which had about 1000 termites). Ta. Fungi were observed growing on most of the termite carcasses in the soil. Ta. M. ant7E was isolated from the carcass recovered from the nest.

At the end of the experiment (67 days after setting up the experiment), the termite nest and The arena's soil and "timber frame" were removed and thoroughly examined. one nest (death (observed extinct termites), less than 10% of the termites are alive. there was.

In other words, more than 90% of the termites in this nest died. died in other nests The number of termites recovered from the nest and soil was not observed and the number of termites recovered from the nest and soil was It was close to the number of li. More importantly, the structure of the wood is subject to termite attack. There was no visible damage caused by the accident.

Control in which the formulation used for construction does not contain M, aniso 1iae In the experiment, termites were observed on the wood structure after one week. Next, termites dug a tunnel in and around the timber. Contains second stage termite adults and eggs. Several hundred termites were observed living inside the timber frame.

The interaction between termites and fungi appears to occur in three stages. First, the colony mem- bers It took one week for the birds to forage over a distance of 120 cm and encounter a structure.

Next, termites in the treated structure recognize the presence of MetarhiZiLlml. be recognized and avoided. When the fungus reached its maximum growth in the soil, it began to die. Avoidance The properties were quickly lost, and the invading termites overcame the obstacle. deer However, freshly prepared conidia were still present in the soil. Shiroa foraging When the termites pass through the area where the fungal preparation has been applied, the conidia come into contact with the termites and infected them. The forager also carries the conidia back to the nest and shares them with other colony members. infection. Its action is apparently fast enough (and the number of termites is moderately large) Even when protecting structures.

Example 13: Prototype M, aniso 1iae ATCC62176 /G.

Soil arena test of a chamber planted with trabe ATCC13021.

Chamber: Q, trabetv sawdust, M, aniso 1iae A Mix TCC62176 formulation and 62176 conidia uniformly in a ratio of 100:5:1. The mixture filled a paper tube (diameter 3 cm x height 5 am).

Blank control chamber: G, trabeum sawdust and grains 1 Paper tubes filled with a ratio of 00:5.

The arena (122X122X30) and termites were the same as in Example 9.

Each vetri dish contained approximately 1000 termites and was treated as a colony.

The arena was filled with soil to a depth of 50 Il.

In each arena, there is one termite colony in the center of the arena. , and there were four infection chambers.

locating each chamber toward and in the center of the wall of each arena; buried in soil. As a repeated test, three arenas were ATCC62176/G, trabeum ATCC13021 chamber Setup and also control one arena as a control channel. I set it up with a camcorder.

In all arenas treated with 62176, one week after the start of the study, Termites were observed in two of the four chambers. However, 2~ After three weeks, there were still termites in the soil, but there were no termites in the chamber. It was not possible to observe a loathe. End the experiment in the control arena 7 From day 30 to day 0, termites were observed in one of the chambers.

62716 to determine termite mortality in these soil arenas. The two prepared arenas were disassembled in 45 days. 30%-40% in both arenas Achieved termite mortality rate. On the 70th, the third chamber 62176 Achieved 100% mortality rate in controlled arena, while in controlled arena We did not observe a decrease in the number of carpenter ants. Furthermore, in the control arena, the second period The presence of termite adults, eggs and early instars was also observed.

From this experiment, L'' 11■ae ATCC62176 and G, trabe Sawdust chamber planted with un ATCC13021 attracts termite population It could be concluded that it could be subsequently eliminated.

F/σ　2σ Time after processing (days) r/c,zD Continuation of front page (81) Designated countries EP (AT, BE, CH, DE.

DK, ES, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE) , 0A (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN.

TD, TG), AU, BB, BG, BR, BY, CA.

CZ, FI, HU, JP, KP, KR, KZ, LK, MG, MN, MW, No. NZ, PL, RO, RU, SD, SK, UA, VN (72) Inventor: Duane, Hongyu United States of America Massachusetts 01002, Amherst Brittany Mainer-115-1

Claims (1)

[Claims] 1. A composition for killing termites, which kills fungi pathogenic to termites. an effective amount of a recruitment promoter to overcome the repellency of the pathogenic fungus to ants; Compositions containing in combination. 2. The recruitment promoter for termites is Gloeophyllum trab. The composition according to claim 1, which is eum. 3. The Gloeophyllum is a fungal culture grown on a nutrient medium. , the composition of claim 2. 4. The recruitment promoter for termites is a chemical produced by wood-decaying fungi. A composition according to claim 1, which is a composition. 5. The composition of claim 1, further comprising a housing for the pathogenic fungus. . 6. The housing may be made of wood materials, cellulose-based products, and synthetic plastics. 7. The composition of claim 6, comprising a material selected from the group consisting of tic. 7. 7. The composition of claim 6, further comprising a coating for the housing. . 8. 8. Composition according to claim 7, consisting of plastic, metal and wood. A composition disposed in a degradation-resistant coating formed of a material selected from the group. 9. 8. The composition of claim 7, wherein the coating is biodegradable. 10. The pathogenic fungi are Metarhizium, Beauveria, E. ntomophthora, Conidiobolus, Paecilomyc selected from the group consisting of es, Verticillium, and Absidia sp. The composition according to claim 1, wherein the composition is 11. The fungi consist of Metarhizium and Beauveria. 11. A composition according to claim 10, selected from the group. 12. A composition according to claim 1, wherein the composition is mixed with a cellulose-based food product. A composition in the form of a preparation containing fungal hyphae. 13. A method of attracting termites, in which G is applied to the site where termites are to be attracted. A method comprising the step of providing loeophyllum trabeum. 14. Claim wherein the Gloeophyllum is provided as a live bacterial culture. The method described in 13. 15. Gloeophyllum is provided as an extract of a fungal culture. The method according to claim 13. 16. pathogenic to termites in combination with the Gloeophyllum 14. The method of claim 13, further comprising providing a fungus. 17. 17. The pathogenic fungus is a termite repellent. the method of. 18 The pathogenic fungi include Hetarhizium, Beauveria, Ve rticillim, Entomophthora, Conidiobolus , Paecilomyces and Absidia spp. 18. The method of claim 17. 19. The pathogenic fungi are Metarhizium anisopliae, and Beauveria bassiana. The method according to item 18. 20. 17. The method of claim 16, further comprising a housing for the fungus. 21. The housing may be made of wood materials, cellulose-based products, and synthetic plastics. 21. The method of claim 20, comprising a material selected from the group consisting of sticks. 22. 21. The method of claim 20, further comprising a covering for the housing. Law. 23. 14. The method of claim 13, wherein the method comprises: locating said pathogenic fungi in combination with an attractant at a site where 14. The method according to claim 13. 24. 14. The method of claim 13, wherein the method comprises: providing a preparation containing fungal hyphae at the site where the structure is to be protected. how to. 25. For those who want to avoid termites in structures that should be protected from termite attack. A method of eliminating pathogenic fungi and repellent fungi at certain predetermined locations of the structure. A method comprising the step of providing a fungus selected from the group consisting of: 26. The fungi include Metarhizium, Beauveria, and Entom. ophthORA, Verticillium, Conidiobolus, P aecilomyces and absidia species. The method according to claim 25-3. 27. 27. The method of claim 26, wherein the fungus is Metarhiziun. 28. 26. The fungus according to claim 25, wherein the fungus is prepared in a formulation for mixing into soil. Method described. 29. 2. The repellent is an extract of a pathogenic or other fungal culture. The method described in 5. 30. further comprising placing the formulation within the chamber of the termite colony. 29. The method of claim 28. 31. The fungal culture is inserted into the soil around the structure to be protected. 26. The method of claim 25, wherein the method is disposed in a housing. 32. A formulation that repels or kills termites and does not cause termites to become fatally infected. Fungi selected from the group of fungi that repel termites and fungi that repel termites are added to the cellulose base. A formulation containing in combination with a composition of fungi, wherein the fungi contain spores, mycelia and and combinations thereof. 33. The fungi include Metarhizium, Beauveria, and Entom. ophthora, Conidiobolus, Paecilomyces and 33. The formulation of claim 32, wherein the formulation is selected from the group consisting of Absidia sp. 34. The fungus is Metarhizium, and the fungus is in the form of a mycelium. 34. The formulation according to claim 33, which is in the form of 35. 34. The formulation of claim 33, wherein the fungus is Beauveria. 36. The fungi screened soil isolates for their effects on termites. 34. The formulation according to claim 33, obtained by 37. The cellulose-based composition consists of bran, grain, and sawdust. 33. A formulation according to claim 32, selected from the group.