Compositions, methods, and devices for controlling and monitoring insects

Abstract

The present invention provides a composition for attracting Carpophilus trunkatus beetles, preferably comprising one or more pheromone compounds produced by male Carpophilus trunkatus beetles. The present invention also relates to kits, methods, and apparatus for attracting, capturing, and monitoring Carpophilus trunkatus beetles.

Description

[Technical Field]

[0001] The present invention relates to devices, compositions, and methods for insect control, and more specifically to a multi-component composition to be used in combination with a device for releasing the composition, and to a device using the composition for attracting, capturing, and / or monitoring insects, more specifically the beetle Carpophilus truncatus. [Background technology]

[0002] Carpophilus trunkatus is a major almond pest, causing significant damage to developing kernels. To control this pest, current "attract and kill" strategies rely on baits consisting of (i) beetle aggregation pheromones (produced by adult male Carpophilus beetles that attack drupes) and (ii) microbially derived synthetic food attractants.

[0003] Carpophyllus trunkatus has recently been identified as a pest of walnuts and other nuts, including pistachios.

[0004] While pheromones and analogues of various Carpophilus species were previously synthesized and identified in the 1990s, the pheromones of Carpophilus truncatus have not been sufficiently studied (Bartelt et al. 1990, 1992; Bartelt 2010). Furthermore, existing lures developed to control Carpophilus beetles that attack drupes are not effective against C. truncatus.

[0005] Common problems in developing control strategies for Carpophilus beetles include the type of trap, trap location, type of food-related odor, pheromones and formulations used in traps, and the lifespan of bait / attractants. Some key attractants may experience a decrease in release rate within 24 hours of bait placement in the field. Furthermore, targeting specific beetle species presents challenges, as generalized compositions fail to achieve the desired specificity, resulting in reduced effectiveness in crop protection (Bartelt 2010).

[0006] Thus, there is a need to overcome, or at least mitigate, one or more of the difficulties or shortcomings associated with the prior art. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] International Patent Application No. PCT / AU2021 / 050240 [Patent Document 2] PCT / AU2021 / 050242 [Overview of the Initiative]

[0008] In one embodiment, the present invention relates to a composition for attracting the beetle Carpophyllus trunkatus, comprising 3,5,7-trimethyl-2,4,6,8-undecatetraene or its geometric isomers, and one or more C6-C6 16 The present invention provides a composition containing an aldehyde.

[0009] As used herein, the term “composition” means a mixture of components, which may be in the form of a mixture of solids, liquids, gases, vapors, gels, or any other suitable phases of the components.

[0010] The geometric isomers of 3,5,7-trimethyl-2,4,6,8-undecatetraene are (2E,4E,6E,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2E,4E,6E,8Z)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2E,4E,6Z,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2E,4Z,6E,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2Z,4E,6E,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2E,4E,6Z,8Z)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2E,4Z,6E,8Z)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2Z,4E,6E,8Z)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2E,4Z,6Z,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2Z,4E,6Z,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2Z,4Z,6E,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2E,4Z,6Z,8Z)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2Z,4E,6Z,8Z)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2Z,4Z,6E,8Z)-3,5,7-trimethyl-2,4,6,8-undecatetraene, (2Z,4Z,6Z,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, The group consisting of (2Z,4Z,6Z,8Z)-3,5,7-trimethyl-2,4,6,8-undecatetraene can be selected.

[0011] In a preferred embodiment, 3,5,7-trimethyl-2,4,6,8-undecatetraene is (E,E,E,E)-3,5,7-trimethyl-2,4,6,8-undecatetraene.

[0012] In some embodiments, the composition does not contain (2E,4E,6E,8E)-7-ethyl-3,5-dimethyl-2,4,6,8-decatetraene.

[0013] In one embodiment, one or more C6-C 16 aldehydes are saturated aldehydes. In another embodiment, one or more C6-C 16 aldehydes are straight-chain aldehydes. In yet another embodiment, one or more C6-C 16 aldehydes are straight-chain saturated aldehydes. In a preferred embodiment, one or more C6-C 16 aldehydes are selected from hexadecanal, pentadecanal, tetradecanal, tridecanal, dodecanal, undecanal, decanal, nonanal, octanal, heptanal, and hexanal, particularly selected from tetradecanal, hexanal, and nonanal, and most particularly selected from tetradecanal.

[0014] In some embodiments, the ratio of 3,5,7-trimethyl-2,4,6,8-undecatetraene to C6-C 16 aldehydes, particularly the ratio of (E,E,E,E)-3,5,7-trimethyl-2,4,6,8-undecatetraene to tetradecanal, is a ratio that results in the release of a ratio of 3,5,7-trimethyl-2,4,6,8-undecatetraene to C6-C 16 aldehydes of 2:1 to 1:5, particularly a ratio of 3,5,7-trimethyl-2,4,6,8-undecatetraene to C6-C 16 aldehydes of 2:1 to 1:2, more particularly a ratio of about 1:1. In some embodiments, the ratio of 3,5,7-trimethyl-2,4,6,8-undecatetraene to C6-C 16 aldehydes in the composition is 2:1 to 1:5, particularly 1:1 to 1:4, more particularly 1:2 to 1:4 or 1:3 to 1:3.5, and most particularly about 1:3.3. A composition containing one of these ratios is 3,5,7-trimethyl-2,4,6,8-undecatetraene and C6-C 16It may result in the release of a 1:1 ratio of aldehydes.

[0015] In one embodiment, the composition described herein further comprises dimethylpyrazine. Thus, in one embodiment, the present invention is a composition for attracting Carpophilus truncatus beetles, comprising 3,5,7-trimethyl-2,4,6,8-undecatetraene or a geometric isomer thereof, and one or more C6-C 16 aldehydes, and a composition comprising dimethylpyrazine, 3,5,7-trimethyl-2,4,6,8-undecatetraene or a geometric isomer thereof, one or more C6-C 16 The aldehydes are as defined above. In a preferred embodiment, the dimethylpyrazine is selected from 2,5-dimethylpyrazine and 2,6-dimethylpyrazine.

[0016] In a preferred embodiment, the composition described herein further comprises an antioxidant. Examples of suitable antioxidants include butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), tocopherol, ascorbic acid, and citric acid. In a particular embodiment, the antioxidant is butylated hydroxytoluene.

[0017] In embodiments, the composition described herein further comprises a carrier. In some embodiments, the carrier can be a solid, semi-solid, gel, or liquid, or a combination thereof. Suitable liquid carriers are volatile solvents, such as volatile nonpolar solvents capable of solubilizing the components of the present composition. In a particular embodiment, the carrier is a nonpolar hydrocarbon. Examples of suitable volatile nonpolar hydrocarbons include pentane, hexane, heptane, and cyclohexane, or mixtures thereof. In a preferred embodiment, the carrier is hexane. In some embodiments, the carrier is used to prepare the composition and load it into a dispenser. In some embodiments, the carrier may be at least partially evaporated from the dispenser before the dispenser is stored or placed in a trap.

[0018] In some embodiments, the composition contains 10 mg / mL of 3,5,7-trimethyl-2,4,6,8-undecatetraene in n-hexane and 5-50 mg / mL of C6-C6. 16 Using an aldehyde and 1 mg / mL of an antioxidant, particularly 10 mg / mL of 3,5,7-trimethyl-2,4,6,8-undecatetraene and 10-50, 20-40, or 30-35 mg / mL of C6-C6 16 It is prepared using an aldehyde and 1 mg / mL of an antioxidant. In a particular embodiment, the composition is an n-hexane composition containing about 10 mg / mL of (E,E,E,E)-3,5,7-trimethyl-2,4,6,8-undecatetraene, about 33.3 mg / mL of tetradecanal, and 1 mg / mL of BHT.

[0019] In a further embodiment, the present invention provides a dispenser containing the composition described herein. In a preferred embodiment, the dispenser allows for the sustained release of the composition described herein.

[0020] In some embodiments, the dispenser includes septums such as rubber septums, beads, granules, pellets, or fragments of resin or inert polymer. In preferred embodiments, the dispenser includes septums.

[0021] In a further embodiment, the present invention provides a composition and / or dispenser described herein, as well as an apparatus including a housing for attracting the beetle Carpophyllus trunkatus.

[0022] As used herein, the term “housing” means any suitable element capable of housing the composition and / or dispenser, which allows for the release of the composition to the surrounding environment and to the outside of the housing. The release of the composition from the housing may be passive or active.

[0023] In some embodiments, the apparatus further includes a container for holding captured Carpophilus beetles, and a lid for the container that allows the beetles to enter the container and prevents them from leaving it. In some embodiments, the container is removable from the lid to allow for the removal of captured beetles. A housing for storing and releasing the composition may be connected to the container or lid to attract beetles to the entrance of the container through the lid. In some embodiments, the lid and housing of the container are green, for example, medium green, medium dark green, or dark green. In some embodiments, the container is transparent or opaque, and is particularly transparent. A preferred apparatus is shown in Figure 7.

[0024] Another exemplary apparatus described herein may include the apparatus described in International Patent Application PCT / AU2021 / 050240, the entire disclosure of which is incorporated herein by reference.

[0025] In some embodiments, the apparatus, One or more co-attractant compounds; and insecticide This further includes one or more of the following.

[0026] In certain embodiments, one or more co-attractant compounds are C1-C6 alcohols, C1-C4 aldehydes, indoles, and C1-C 12 The co-attractant compounds may be selected from esters. In preferred embodiments, one or more co-attractant compounds may be selected from the group consisting of ethanol, isopentyl alcohol, acetaldehyde, isobutanol, 2-methylbutanol, ethyl acetate, isopentyl acetate, isobutyl acetate, 2-phenylethyl acetate, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), methyl benzoate, and (Z)-3-hexenyl acetate. In specific embodiments, one or more co-attractant compounds include ethanol and isopentyl alcohol, and in particular aqueous ethanol and isopentyl alcohol.

[0027] As used herein, the term “aqueous” means a water-based solvent, preferably containing at least about 40% water, particularly distilled water, and may also contain other water-soluble or water-miscible components.

[0028] In some embodiments, the aqueous ethanol is 30% to 60% ethanol in water, particularly 40% to 55% ethanol in water, and more particularly 40% to 50% ethanol in water, for example, 45% ethanol in water.

[0029] In one embodiment, the apparatus may include one or more co-attracting compounds, each of which is located in a separate container or deposit element.

[0030] As used herein, a deposit element means any suitable object from which a compound can be stored and from which a compound can be released. In embodiments, the deposit element may be a cotton roll / dental wick, or any other object suitable for storing and releasing co-attracting compounds.

[0031] In some embodiments, a deposit element containing a co-attractant compound is placed in a container that releases the co-attractant compound at a desired rate. In preferred embodiments, the container is made of low-density polyethylene. In one embodiment, the thickness of the low-density polyethylene allows for control of the co-attractant compound release rate. Preferably, the container is made of low-density polyethylene having a thickness of approximately 25 μm to 250 μm, more preferably approximately 35 μm to 225 μm. In particularly preferred embodiments, the container is made of low-density polyethylene having a thickness of approximately 50 μm to 200 μm.

[0032] In alternative embodiments, the apparatus may include one or more co-attractant compounds, which are present in a co-attractant mixture.

[0033] In a preferred embodiment, the apparatus may contain one or more of the above-mentioned co-attracting compounds present in the co-attracting compound mixture, the mixture being in the form of a solid, liquid, gas, vapor, gel, or any other suitable phase, preferably in the form of a liquid or gel. In one embodiment, the apparatus may contain one or more co-attracting compounds present in the mixture, the mixture being in the form of a solution, the co-attracting compound mixture containing ethanol and isopentyl alcohol, particularly aqueous ethanol and isopentyl alcohol. In another embodiment, the apparatus may contain one or more co-attracting compounds present in the mixture, the mixture being in the form of a gel, the co-attracting compound mixture containing aqueous ethanol and isopentyl alcohol. Suitable gel-forming components include carbomer, glycerin, and tertiary amines such as triisopropanol.

[0034] In one embodiment, the insecticide is an organophosphate. In a preferred embodiment, the insecticide is selected from the group consisting of dichlorvos, thiometon, naled, parathion, malathion, and S-benzyldiisopropylphosphorothiolate (IBP), and is particularly selected from dichlorvos.

[0035] In some embodiments, the device or dispenser regulates the release of the composition. In a preferred embodiment, the device or dispenser regulates the release of the composition for approximately 1 to 8 weeks. In a more preferred embodiment, the device or dispenser regulates the release of the composition for approximately 2 to 8 weeks. In a more preferred embodiment, the device and / or dispenser regulates the release of the composition for approximately 4 to 8 weeks.

[0036] In some embodiments, the device regulates the release of a co-attractant compound or co-attractant mixture. In preferred embodiments, the device regulates the release of the co-attractant compound or co-attractant mixture for approximately 1 to 8 weeks. In more preferred embodiments, the device regulates the release of the co-attractant compound or co-attractant mixture for approximately 2 to 8 weeks. In even more preferred embodiments, the device regulates the release of the co-attractant compound or co-attractant mixture for approximately 4 to 8 weeks.

[0037] In some embodiments, the apparatus replaces one or more of the composition, co-attractant compound or mixture, and insecticide. These components may be replaced when the regulated release becomes insufficient, for example, when they are no longer effective in attracting and / or killing Carpophyllus truncatus beetles.

[0038] In further embodiments, the present invention provides a kit comprising a composition and / or dispenser described herein, and a capture device. In some embodiments, the kit comprises a composition or dispenser described herein, and the capture device is an apparatus comprising a housing and a container with a lid described herein. In preferred embodiments, the kit comprises a dispenser and a capture device described herein. In preferred embodiments, the components of the kit may be assembled so that, in normal use, the dispenser allows for the slow release of the composition described herein, and the capture device captures Carpophyllus trunkatus beetles. In another preferred embodiment, the components of the kit may be assembled so that the capture device and composition are suitable for or can be used in the method of the present invention described herein.

[0039] In certain embodiments, the kit is One or more co-attractant compounds described herein; and insecticides described herein This further includes one or more of the following.

[0040] In some embodiments, the kit may include multiple compositions of the present invention, multiple co-attractant mixtures or multiple sets of co-attractants housed in separate containers, and / or multiple insecticide components, allowing for the replacement of these components within the capture device at intervals over the growth / harvest or monitoring period, for example, when the effectiveness of the composition, co-attractants, and / or insecticide activity decreases.

[0041] In a further aspect of the present invention, a method for attracting and / or capturing the beetle Carpophyllus trunkatus is provided, comprising the step of exposing an environment infested with the beetle to a composition, dispenser and / or apparatus described herein.

[0042] In some embodiments, the environment in which the disease is prevalent is an orchard, more specifically a nut orchard, in particular an almond orchard, a pistachio orchard, a walnut orchard, a cashew orchard, a kumili orchard, a macadamia orchard, and / or a Brazil nut orchard. In certain embodiments, the orchard may be an almond orchard, a pistachio orchard, or a walnut orchard.

[0043] In other embodiments, the permeated environment is a stockpile of nuts, particularly stockpiles of almonds, pistachios, walnuts, cashews, kumiri nuts, macadamia nuts, and / or Brazil nuts. In certain embodiments, the nut stockpile may be a stockpile of almonds, pistachios, or walnuts.

[0044] In certain embodiments, the environment in which the disease is prevalent is an almond orchard or an almond stockpile, and in particular an almond orchard.

[0045] When used in nut orchards such as almond orchards infested with C. trunkatus beetles, the composition is optionally placed in the trapping device and is preferably installed in the orchard at intervals of 10m to 100m, particularly 20m to 80m, more particularly 30m to 70m, and even more particularly 40m to 60m, for example, at 50m intervals. In some embodiments, the trap density is 3 to 25 traps per hectare, particularly 10 to 20 traps per hectare, for example 15 to 17 traps per hectare.

[0046] A further aspect of the present invention provides a method for monitoring the presence of the beetle Carpophyllus trunkatus, comprising the step of placing a composition, dispenser, and / or apparatus described herein into an environment in which the presence of the beetle needs to be monitored.

[0047] In certain embodiments, the environment in which the presence of Carpophilus trunkatus beetles needs to be monitored is a nut orchard, a nut stockpile, or nuts ready for export or recently imported. The nuts can be selected from almonds, pistachios, walnuts, cashews, kumiri nuts, macadamia nuts, and Brazil nuts, and in particular from almonds, pistachios, and walnuts. In certain embodiments, this environment is an almond orchard, an almond stockpile, or almonds ready for export or recently imported.

[0048] In some embodiments, the composition and attractant, and optionally the insecticide, may be replaced at regular intervals over the spread or monitoring period. For example, the composition and / or co-attractant mixture may be replaced occasionally 1 to 8 weeks after the trap is placed, and thereafter every 1 to 8 weeks over the growing / harvesting or monitoring period. For example, the composition and / or co-attractant mixture may be replaced every 2 to 8 weeks or every 4 to 8 weeks. In some embodiments, the insecticide may be replaced at the same time as the composition and / or co-attractant mixture.

[0049] In this specification, the term "including" and its variations are not intended to exclude the existence of other integers, components, or steps.

[0050] In this specification, no reference to prior art in this specification shall be deemed, nor shall it be construed, as an agreement or any suggestion in any form that such prior art forms part of the common general knowledge in Australia or any other jurisdiction, or that such prior art can be reasonably expected to be combined by those skilled in the art.

[0051] The present invention will be described in more detail with reference to the attached examples and drawings. However, it should be understood that the following description is merely illustrative and should not be considered in any way to limit the generality of the invention described above. [Brief explanation of the drawing]

[0052] [Figure 1] This figure shows the setup used for dynamic headspace sampling and pheromone collection. [Figure 2] This figure shows GC-MS chromatograms of almond kernels infested with either male (♂) or female (♀) beetles. Hexanal and dimethylpyrazine are compounds associated with beetle infestation, regardless of beetle sex (and are absent in kernels without beetles). The remaining shaded compounds are characteristic of male beetle infestation. [Figure 3a-e]This figure shows GC-MS chromatograms of odors from living beetles and synthetic pheromones (gray). A comparison of retention indices of natural and synthetic pheromones was used to confirm compound IDs and reliability. a) corresponds to the odor of male beetles fed artificial feed. b) is a chromatogram of the odor of male beetles fed almond kernels. c) is a chromatogram of synthetic pheromone 2 supplied by Boron Molecular (Melbourne). d) corresponds to a chromatogram of a synthetic male-specific compound (newly identified pheromone, commercially available). e) shows the overlays of Figures 3a-3d. The gray overlays match the retention times of individual synthetic pheromones to the retention times of natural pheromones found in live beetle extracts, and are Phero#2 (pheromone 2) and Phero#3 (presumed pheromone). [Figure 4] This figure shows an SPME-GC-MS chromatogram representing the odor blend generated by septums loaded with Phero#2 and Phero#3 (tetradecanal) and an antioxidant (butylated hydroxytoluene) prior to field trials. [Figure 5] This figure shows a map of the orchard plots used for field evaluation of pheromone septums. The numbers indicate the location of individual traps within the orchard. The arrows indicate different plots used in the randomized complete plot design (10 plots = 10 replicates, each containing 6 test treatments). [Figure 6] This figure shows bar graphs representing the beetle collection rates in field trials using different pheromone lures in combination with a standardized co-attractant blend. Gray bars represent the collection rate of C. trunkatus, and white bars represent the number of other Carpophilus beetles (mainly C. hemipterus) collected in the traps. Error bars represent the standard error. Lowercase lettering above the gray bars indicates the statistical difference between the collection rates of C. trunkatus beetles for different treatments, and uppercase lettering above the white bars indicates the statistical difference for other Carpophilus species. Each treatment was repeated 10 times, and beetle samples were collected once every two weeks. [Figure 7]Figure 7a shows a typical capture device for use with the composition of the present invention; Figure 7b shows the assembled state; and Figure 7b shows the disassembled state. [Figure 8] This figure shows a bar graph illustrating the average number of C. trunkatus beetles (gray bars) and other Carpophyllus species (white bars) collected during a two-week trial. Error bars represent the inversely transformed 95% confidence level. Uppercase and lowercase lettering indicate statistically significant differences in collection amounts for C. trunkatus and other Carpophyllus species using different pheromone treatments. [Figure 9] This figure shows a bar graph indicating the number of larvae produced per three adult females for each nut food source used. [Figure 10] This figure shows a bar graph indicating the number of larvae that reached the final instar per cycle for each nut food source used. [Figure 11] This figure shows the trap layout for the mass capture plot. X represents a trap, and the shaded areas represent sampling subplots; NP represents a nonpareil row. [Figure 12] This is a photograph showing a trap fixed in place within an orchard. [Figure 13] This figure shows a bar graph illustrating the total number of C. truncatus captured per plot over a two-week period. [Figure 14] This figure shows a bar graph illustrating the average percentage of damage dealt by C. truncatus under control and mass trapping conditions. Error bars indicate SEM. [Modes for carrying out the invention]

[0053] Example 1 - Specimen and pheromone collection i. Plants and insects The raw almonds used in all experiments were collected from a commercial almond orchard located near Mildura, Victoria, Australia. The Carpophilus beetles (Carpophilus trunkatus) used to infest the almonds were obtained from a laboratory colony maintained at the AgriBio Centre for AgriBiosciences (Bundoora, Australia). This colony was built from wild-collected beetles from the same orchard. Newly emerged beetles were collected and their sex was determined under a stereomicroscope before infesting the nuts.

[0054] ii. Chemical substances The previously identified C. trunkatus pheromone compound, pheromone 2:(E,E,E,E)-3,5,7-trimethyl-2,4,6,8-undecatetraene (disclosed in PCT / AU2021 / 050242 above), was synthesized by Boron Molecular (Noble Park, VIC, Australia). The newly identified pheromone compound, tetradecanal, was purchased from Ambeed Inc. (USA) through its regional distributor (Sigma Aldrich Australia). Dichloromethane, ethanol (96% purity), nonyl acetate, butylated hydroxytoluene, and isopentyl alcohol were purchased from Sigma-Aldrich (Castle Hill, NSW, Australia).

[0055] iii. Collection of volatile substances Volatile substances from beetle-free (non-infested) and infested kernels were collected by dynamic headspace sampling in the apparatus shown in Figure 1. Three treatments were prepared for odor sampling: (i) male beetles, (ii) female beetles, and (iii) no beetles. For each treatment group, 25 kernels (15 whole and 10 cut in half) were placed in a 300 mL glass container used for volatile substance collection. Treatments containing beetles (i and ii) included 60 adult beetles of the required sex. The beetles were fed kernels for one week prior to sampling. Volatile substance collection was performed by circulating airflow through two glass sockets located on the opposite side of the glass container, which were used as intake and exhaust ports. Purified air drawn through an activated carbon filter attached to the intake port (outside) circulated the volatile substances in the chamber at a set flow rate (100 mL / min). -1 The samples were then transported together to an adsorbent filter connected to the exhaust port (the vacuumed interior). The collection container was wrapped in aluminum foil to create a dark environment preferred by beetles. Multiple collections were performed simultaneously using a six-arm manifold made of PVC tubing connected to a single vacuum point (shown in Figure 1). Small valves were used to adjust the airflow on different arms of the manifold, and this was controlled using an anemometer. The adsorbent filter used to capture volatile substances consisted of 100 mg of Porapak Q powder packed between two silane-coated glass wool plugs inside a glass Pasteur pipette. Volatile substances were collected from 12 samples for each treatment over a period of 7 days. At the end of the collection, 2 mL of dichloromethane was used to elute the volatile substances from the adsorbent filter. After adding 500 ng of nonyl acetate to the sample as an internal standard (IS), these were condensed to a final volume of approximately 100 μL by solvent evaporation under a gentle flow of nitrogen.

[0056] Example 2 - Chemical analysis of pheromone samples Next, the pheromone samples obtained according to Example 1 were used for chemical analysis and field experiments.

[0057] Volatile substance samples dissolved in dichloromethane were analyzed by gas chromatography-mass chromatography (GC-MS). An Agilent 7890B gas chromatograph, integrated with a single quadrupole Agilent 5977B mass spectrometer and equipped with an Ultra Inert HP-5MS capillary column (30 m × 0.25 mm × 0.25 μm), was used with an Agilent 7650 ALS autosampler. 2 μL aliquots were injected at 250°C in spitless mode. The initial oven temperature was set to 40°C and held for 2 minutes, then increased to 220°C at a rate of 10°C / min. -1 Then increase the temperature by 20°C per minute until the final temperature reaches 300°C. -1 The temperature was increased and maintained for 1 minute. Mass spectra were acquired in EI mode (70 eV) with the mass scanning range set to 35-550. The quadrupole and ionization source temperatures were set to 150°C and 230°C, respectively. Compounds were tentatively identified using the NIST 14 mass spectrum library, either by comparing its Kovats index to the index of directly available literature or by injecting commercially available synthetic compounds.

[0058] Data Analysis Chromatographic data from GC-MS analysis of volatile substances in almonds and beetles were extracted using the built-in deconvolution and peak alignment tools from the eRah package in R studio (Domingo-Almenara et al. 2016). The amounts of different compounds were estimated in ng IS equivalent / week by comparing the extracted peak area to the peak area of ​​an internal standard. The differences between the obtained estimators in the headspace of kernels infested with control, male, and female were then tested using ANOSIM (analysis of similarity) with the Bray-Curtis dissimilarity matrix. For multi-level pattern analysis, the indicspecies package (De Caceres et al. 2011) was used to investigate the compound characteristics of different odor profiles.

[0059] Results - Analysis of volatile substances from infested almond kernels and beetle-free almond kernels. GC-MS analysis of odor extracts (Figure 2) demonstrates differences between the chemical profiles of almond kernels free from beetles and those infested with beetles. Dimethylpyrazine compounds (2,5- or 2,6) were almost absent in the headspace of beetle-free kernels and present in infested samples regardless of beetle sex. Elevated hexanal concentrations were also observed in the odor of kernels infested with both sexes of beetles. Using multi-level pattern analysis, four compounds were identified as specific to male beetle infestation.

[0060] Among these compounds were (2E,4E,6E,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene (pheromone 2 or Phero#2), previously identified as the main C. trunkatus pheromone (identified in its entirety in PCT application PCT / AU2021 / 050242, which is incorporated herein by reference), an unidentified compound (mw=218), and a compound identified with high confidence as tetradecanal by mass spectral library. A weak but significant increase in nonanal was associated with the spread of male beetles. The results of GC-MS analysis and the significance of the statistical association with different profiles are presented in Table 1.

[0061] Figure 2 shows GC-MS chromatograms of almond kernels infested with either male (♂) or female (♀) beetles.

[0062] [Table 1A]

[0063] [Table 1B]

[0064] Matching compound IDs using synthetic products The reliability and purity of the compound were verified by GC-MS analysis by comparing the mass spectrum and retention index of the synthesized compound with those obtained from the odor extract of beetles that had eaten almond kernels (see Figures 3a-e).

[0065] The mass spectra of the synthetic compounds matched those of their natural counterparts, and their retention indices were close enough to infer their reliability (Pheromone 2 [(2E,4E,6E,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene]: 1486 vs. 1489, Pheromone 3 [Tetradecanal]: 1616 vs. 1613). The purity of the newly prepared synthetic pheromone (Pheromone 2) was 86%, and the purity of Pheromone 3 (Tetradecanal) was approximately 97%. The amounts of the two compounds to be loaded into rubber septums were adjusted using SPME-GC-MS analysis so that the compounds would be released in approximately equal ratios. The results of this analysis are shown in Figure 4.

[0066] Example 3 - Field experiment of pheromone samples Field experiments were designed to test the attractiveness of C. trunkatus pheromone 2, previously described, under field conditions, and the pheromone (attractant) activity of the newly identified pheromone 3. The pheromones were tested individually or in combinations equivalent to those found in biological samples. The tests included commercially available pheromone mixtures developed for the control of drupe Carpophilus species ("Tri-species lure, Catcha® pheromone lure, (Insect Management Services, Baccus Marsh, VIC, Australia)"). All pheromones were tested with co-attractant mixtures in the form of aqueous ethanol and isopentyl alcohol solutions that synergistically act in C. trunkatus to elicit a strong behavioral response.

[0067] Pheromone test lures were loaded onto white Precision Seal® rubber septums (8 mm OD, Sigma Aldrich product code: Z553913), and butylated hydroxytoluene was used as an antioxidant. The amount of each compound applied to the septum was determined by SPME-GC-MS analysis of the test septums on days 1 and 3, so that the two pheromones were produced in approximately equal ratios. The amount of antioxidant was selected as 10% of the main pheromone, based on existing literature (Table 2). Hexane dilutions of the neat pheromones were prepared, and the desired amount was loaded onto the septums. The loaded septums were dried overnight under a fume hood and stored at -20°C in heat-sealed foil bags until use in field trials.

[0068] [Table 2]

[0069] Field trials were conducted from December 2021 in a commercial almond orchard located near Mildura, Victoria, Australia. Orchard plots were selected based on the presence of C. trunkatus populations in guard traps and on nuts on the ground, as well as their size (large enough to accommodate the trials). Pheromone septums were suspended using paper clips inside black bucket traps (Caprophilus Catcha trap, Bugs for Bugs, Toowoomba, QLD) containing 250 mL of a co-attractant solution optimized for capturing C. trunkatus (the entire solution of which is incorporated herein by reference PCT / AU2021 / 050242) and insecticide flakes (Killmaster, dichlorvos, 15 mm × 15 mm). The traps were placed at intervals of 50 m or more, and the treatment was carried out in a randomized, fully partitioned design (each partitioned area corresponding to the crossing of 6 traps) and repeated 10 times (as shown in Figure 5). The lure septum and co-attractant solution were replaced every two weeks, simultaneously with the collection of beetle samples. Beetles present in different traps were identified and counted under a stereomicroscope. The experiment was conducted for a total of 6 weeks.

[0070] Data Analysis Field data were analyzed using a generalized linear mixed model (GLMM) (Brooks et al., 2019) fitted with a negative binomial distribution using the glmmTMB package. The number of C. trunkatus and other Carpophyllus beetles collected was used as the response variable, and "bait treatment" was set as a fixed factor. Other random variables, such as the "date" of evaluation, which constitutes a temporary variable of the beetle population during the experiment, as well as "plot" and "column" which control the heterogeneous spatial distribution of beetles within the plot, were used as random factors when they contributed to improving the fit. The lsmeans package (Lenth 2016) was used for multiple comparison tests (adjusted Tukey post-hoc tests).

[0071] Attraction of previously identified pheromones and newly identified putative pheromones in field trials The best model fit for C. trunkatus beetle collection data was achieved by using treatment as a fixed factor and "date" and "section" as random variables (due to the gradient of the beetle population across sections). The number of C. trunkatus collected varied significantly depending on the pheromone mixture loaded into the septum (χ²). 2 (=822, df=5, p<0.001). The results of this study are shown in Figure 6.

[0072] Septums loaded with pheromones 2 and 3 were observed to collect significantly more C. truncatus beetles than all other treatments (p<0.001). Furthermore, septums containing only pheromone 2 collected significantly more beetles than commercially available septums containing all three pheromones (p<0.001).

[0073] The collection volumes of other Carpophyllus species (C. hemipterus making up the majority) were analyzed using a similar model, adding "columns" as a random variable, taking into account the observation of higher collection volumes at the edge of orchards. There were no significant differences in the amount of beetles collected under the treatment.

[0074] Example 4 - Field experiment of pheromone samples A short trial was conducted over two weeks. Treatment was randomized and repeated in a row of 10 trees with traps spaced approximately 50m apart (10 rows x 2 treatments - 20 repetitions per treatment; 3 treatments for 20 repetitions, using 60 traps). The treatments are shown in Table 3.

[0075] [Table 3]

[0076] The data were analyzed using GLMM in the same manner as in Example 3, with trap collection as the response variable, lure treatment as a fixed factor, and columns and rows as random (spatial features). Tukey's adjusted post-hoc test was used for multiple comparisons of the three families of estimates. The results for the mixed model were obtained using Wald χ². 2 =62.5, df=1, p=2.64 10 -14 Multiple comparisons are shown in Table 4.

[0077] [Table 4]

[0078] Figure 8 shows that Phero 3 collected significantly fewer C. trunkatus and other Carpophilus species than Phero 2 and Phero 2 + Phero 3, suggesting that Phero 3 acts synergistically with Phero 2. However, Phero 2 + Phero 3 collected more C. trunkatus than Phero 2, although the difference was not statistically significant. These results differ from those of Example 3, where Phero 2 and 3 were the most effective treatments by a considerable margin. The beetle population in the orchard was considerably larger in Example 4 (reflecting a larger number of traps across the treatments), which likely influenced the amount of beetles collected in the traps across the treatments. For example, in fruit flies, field trials conducted under high pest pressure with artificial bait formulations showed a similar lack of statistical significance between artificial bait treatments compared to those under low pest pressure (Henneken et al., 2022; Cunningham et al., 2018). These results highlight the likelihood that artificial bait technology is most effective under lower pest pressure.

[0079] Example 5 - Nut Feeding Test A laboratory colony of Carpophyllus trunkatus was constructed by collecting mammy nuts from a commercial almond orchard in the Sunrasia growing area of ​​Victoria, Australia. The insects were reared at 25°C, 12 hours day and night, and 60% relative humidity on a sugar-soybean diet. The colony was maintained at the AgriBio Centre for AgriBioscience, Bundoora, Victoria, and all beetles used in the experiment were obtained from this laboratory colony.

[0080] To determine the basic host range of C. trunkatus in nut-like substrates, nine nut or seed products, encompassing both commercial crops and native plant species, were selected for host compatibility testing based on known or potential uses by C. trunkatus. In both adult survival and larval development tests, 10 replicates were prepared for each nut species by roughly chopping 1.5 g of nuts to expose the interior for beetle access. The chopped nuts were placed in 5 mL plastic specimen tubes, secured with a muslin cover and a screw-top lid with a 3.5 mm hole, maintaining a moist microclimate while allowing airflow. In the control group, equivolume of chopped mounting polystyrene was used to provide a similar microhabitat and maintain moisture, but without access to edible resources, in order to establish baseline parameters of life trait.

[0081] Adult survival and tolerability Six newly emerged adult Carpophyllus beetles, three males and three females, were placed in each tube, and their survival and the development of F1 generation larvae were monitored daily. The number of beetles was selected to avoid premature death, particularly during the first few days of the experiment, because this species aggregates and is less active when isolated and unfed, especially when exposed to the nut substrate. All tubes were misted daily with water to maintain sufficient humidity. The date of death for all adults, as well as the number of fifth-instar larvae born in each tube over the duration of the experiment (100 days), were recorded. If the nuts became excessively wet due to the activity of many F1 larvae, or if the product was nearly depleted, the adults were transferred to new tubes of chopped nuts. Upon completion of the experiment, all adults and larvae were removed and stored in 100% ethanol. A portion of the F1 larvae from each treatment were reared to adulthood to confirm that the adults were fertile and free from abnormalities. The average number of larvae born is shown in Figure 9.

[0082] Larval development Six newly laid C. trunkatus eggs were carefully transferred onto chopped nuts in each tube using a sterile, damp brush. The tubes were monitored daily, and the date when the fifth instar larvae developed and were ready to pupate was recorded. The tubes continued to be monitored for one month after the final larva was observed, at which point it was assumed that any remaining larvae would have died. Some insects from each treatment were reared to adulthood to ensure that the adults were reproductively capable and free from abnormalities. The number of days to reach the fifth instar larva is shown in Figure 10.

[0083] Example 6 - Field experiment to attract and kill insects the purpose By using an attractant to capture large numbers of C. truncatus, we can determine the extent to which the damage to almond kernels can be mitigated.

[0084] Materials and methods Test location This experiment was conducted in a mature, commercial almond orchard in the Robinvale district of Victoria. Fifteen orchard plots potentially suitable for the experiment were identified based on their size and the level of Carpophyllus kernel damage recorded in previous harvests. These plots contained 16-17 year old trees on two rows of drip irrigation, with tree and row spacings of 4.65m and 7.25m, respectively.

[0085] The presence of Carpophyllus infestations in mammy nuts on the ground in these 15 plots was assessed in mid-September 2023. Assessments were performed every 6 rows (43.5 m) for every 10th (46.5 m) or 15th (69.75 m) tree. At each assessment point, up to 10 nuts were opened and examined for live Carpophyllus, and the examination was stopped immediately if beetles were found. Based on this assessment, 10 plots were selected for the trial based on their relatively wide distribution of live Carpophyllus.

[0086] Experimental Design This study employed a randomized plot design using balanced pairs. Ten orchard plots selected for the study were paired based on having similar Carpophyllus damage levels recorded in previous harvests. Two treatments, "mass trap" and "control," were randomly assigned to each pair of plots, and five iterations were prepared for each treatment.

[0087] High levels of kernel damage at harvest had previously been observed in trees up to 90m from areas of mammy nuts that had been infested with high levels of C. trunkatus during the winter. Based on that observation, a plot size of 5.67ha (1,683 trees) was used in this current trial to minimize peripheral effects. This allowed for a capture zone of at least 100m around a “sampling subplot” of 35 trees, including 21 nonpareil trees that would be used to collect nut samples for damage assessment. The same plot / subplot layout was applied to the control plot.

[0088] As shown in Figure 11, 94 traps were installed in each of the large-scale trap plots, resulting in a density of 16.6 traps / ha.

[0089] Trap components and maintenance All traps used in this test were identical and consisted of the following: • A bucket / funnel trap (26.5 x 31 x 41 cm) consisting of a transparent bucket with a green rain cap and funnel, commercially available as the "Carpophilus Angler Trap" (GroChem, Melbourne, Australia). Each trap was secured to the ground surface using custom-made metal stakes and rings, as shown in Figure 12, and held in place with rubber tree ropes. A rubber pheromone septum (Precision Seal® white rubber septum) loaded with (2E,4E,6E,8E)-3,5,7-trimethyl-2,4,6,8-undecatetraene (3 mg), tetradecanal (10 mg), and BHT (0.3 mg). A 250 mL co-attractant solution containing 45% aqueous ethanol and isopentyl alcohol (800 μL / 100 ml) in a plastic container covered with fine gauze to prevent beetles from entering the container. A 1cm x 1cm piece of Killmaster insecticide (dichlorvos) (Barmac Industries Pty. Ltd., Queensland, Australia) for killing beetles caught in traps.

[0090] A fresh co-attractant solution was prepared in advance and stored at 4°C until needed. The co-attractant and pheromone septum were replaced every two weeks, while the insecticide fragments were replaced once a month.

[0091] Timeline The trapping equipment was set up in late September / early October, and the traps were first fitted with pheromone septums, co-attractants, and insecticide fragments on October 10th and 18th. The traps in each experimental plot were maintained until nut samples were collected from that plot, with the earliest sample collection being 15 / 2 / 2024 and the latest being 1 / 3 / 2024.

[0092] Data collection Trap harvesting amount Trap samples were collected every two weeks. Insects collected from each of the five trapping plots were pooled into one sample per plot, sealed in labeled plastic ziplock bags, and temporarily stored at 4°C. After removing all specimens that were not of the genus Carpophyllus, the samples were stored at -20°C and then sent to the AgriBio Centre (Bundoora, Victoria) of Agriculture Victoria for identification. All samples were measured by volume using graduated cylinders.

[0093] For each sampling date, 2 mL of secondary samples were classified from each capture plot, and the species was identified using morphological clues and stereomicroscopy. Then, the total beetle volume was converted to the total number of C. trunkatus captured per plot using C. trunkatus count / mL.

[0094] The sex ratio of captured C. trunkatus was evaluated for 15 samples collected over six sampling dates between early December 2023 and early February 2024. For this, 20 beetles were randomly selected from the 2 mL subsamples of C. trunkatus classified above, and their sexes were determined under a stereomicroscope.

[0095] Jin damage For nut sampling, 10 trees were randomly selected from 21 nonpareil trees in each sampling subplot. Immediately after shaking the trees for commercial harvesting, 100 freshly harvested nuts were collected from the ground beneath each of the 10 selected trees. All tree-only samples were stored in coarse mesh onion bags at approximately 4°C to continue drying and prevent further kernel damage from Carpophilus beetles or carob moths. The presence of kernel damage by Carpophilus beetles was assessed by manually cracking the nuts and, if necessary, visually inspecting them using a Maggylamp or stereomicroscope.

[0096] Data Analysis All statistical tests were performed using GenStat (VSN International, 2023). Restricted maximum likelihood model (REML) analysis was performed, including plot ID as a random effect, to determine whether the treatment could explain the variability at the observed levels of insect damage.

[0097] Results and Discussion Trap harvesting amount The beetle samples collected from the traps by February 1st had been processed, and the total number collected up to that date was just under 684,000 C. truncatus (Figure 13).

[0098] On average, C. trunkatus accounted for 96.4% of the total beetle collection, highlighting the specificity of the lure for this species. Interestingly, the sex ratio of C. trunkatus in the 15 samples was 68% female on average. This bias towards females adds value to the capture program by increasing the impact that capture has on the reproductive capacity of the C. trunkatus population.

[0099] Jin damage The REML model demonstrated a significant treatment effect on the level of nematode damage. Significantly less damage was observed in the mass trapping treatment compared to the control (Figure 14) (df=1; F=6.78, p=0.031). The plot ID contributed only 0.26% to the observed variability in insect damage, suggesting that the plot pairs were well-balanced. Table 5 shows the mean percentage of nematode damage and the reduction in damage due to mass trapping for each plot pair.

[0100] [Table 5]

[0101] This initial attractant and insecticidal trial (A&K trial) demonstrates the potential of the lure for mass capture.

[0102] An exception in these data is that the percentage of kernel damage reduction in plot vs. 4 (18.8%) is lower than the average percentage of reduction across the other four plot pairs (66%). According to the nut evaluators, the sample from the control plot in plot vs. 4 contained a significant number of nuts with tightly sealed shells, and it was suggested that such nuts were less susceptible to kernel damage by Carpophyllus species, thus distorting the results.

[0103] conclusion The average and maximum reductions in damage, 56% and 79%, achieved through the extensive "attracting and killing" treatments used in this experiment, are very promising. The fact that damage was kept below 2% of the unofficial industry threshold in two of the five capture plots is also promising.

[0104] The improved co-attractant and novel pheromone blend demonstrated high selectivity for C. trunkatus, which accounted for over 96% of the total Carpophyllus beetle collection. As a result, this trap and lure can be used by producers as a reliable monitoring tool for this species.

[0105] Finally, it should be understood that various changes, modifications, and / or additions may be made without departing from the spirit of the invention outlined herein.

[0106] References Bartelt RJ (2010) Volatile hydrocarbon pheromones from beetles. In: Blomquist GJ, Bagnieres AG (eds) Insect Hydrocarbons: Biology, Biochemistry, and Chemical Ecology. Cambridge University Press, pp 448-476 Bartelt RJ, Dowd PF, Plattner RD, Weisleder D (1990) Aggregation pheromone of driedfruit beetle, Carpophilus hemipterus Wind-tunnel bioassay and identification of two novel tetraene hydrocarbons. J Chem Ecol 16:1015-1039. doi: 10.1007 / BF01021008 Bartelt RJ, Weisleder D, Dowd PF, Plattner RD (1992) Male-specific tetraene and triene hydrocarbons of Carpophilus hemipterus: Structure and pheromonal activity. J Chem Ecol 18:379-402. doi: 10.1007 / BF00994239 Brooks ME, Kristensen K, Darrigo MR, et al (2019) Statistical modeling of patterns in annual reproductive rates. Ecology 100:e02706. https: / / doi.org / 10.1002 / ecy.2706 De Caceres M, Sol D, Lapiedra O, Legendre P (2011) A framework for estimating niche metrics using the resemblance between qualitative resources. Oikos 120:1341-1350. https: / / doi.org / 10.1111 / j.1600-0706.2011.19679.x Domingo-Almenara X, Brezmes J, Vinaixa M, et al (2016) ERah: A Computational Tool Integrating Spectral Deconvolution and Alignment with Quantification and Identification of Metabolites in GC / MS-Based Metabolomics. Anal Chem 88:9821-9829. https: / / doi.org / 10.1021 / acs.analchem.6b02927 Lenth RV (2016) Least-Squares Means: The R Package lsmeans. J Stat Softw 69:1-33. https: / / doi.org / 10.18637 / jss.v069.i01

Claims

1. A composition for attracting the beetle Carpophyllus trunkatus, comprising 3,5,7-trimethyl-2,4,6,8-undecatetraene or its geometric isomers, and one or more C 6 ~C 16 A composition containing an aldehyde.

2. The composition according to claim 1, wherein 3,5,7-trimethyl-2,4,6,8-undecatetraene is (E,E,E,E)-3,5,7-trimethyl-2,4,6,8-undecatetraene.

3. One or more C 6 ~C 16 The composition according to claim 1 or 2, wherein the aldehyde is a saturated aldehyde.

4. One or more C 6 ~C 16 The composition according to claim 3, wherein the aldehyde is selected from tetradecanal, hexanal, and nonanal.

5. One or more C 6 ~C 16 The composition according to claim 4, wherein the aldehyde is tetradecanal.

6. The composition according to any one of claims 1 to 5, further comprising dimethylpyrazine.

7. The composition according to any one of claims 1 to 6, further comprising an antioxidant.

8. The composition according to claim 7, wherein the antioxidant is selected from butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), tocopherol, ascorbic acid, and citric acid.

9. The composition according to any one of claims 1 to 8, further comprising a carrier.

10. The composition according to claim 9, wherein the support is a nonpolar hydrocarbon.

11. A dispenser comprising the composition according to any one of claims 1 to 10.

12. The dispenser according to claim 11, which enables the sustained release of the composition according to any one of claims 1 to 10.

13. The dispenser according to claim 12, comprising beads, granules, pellets, or fragments of a septum, resin, or inert polymer.

14. The dispenser according to claim 13, which includes a septum.

15. An apparatus comprising a composition according to any one of claims 1 to 10 or a dispenser according to any one of claims 11 to 14, and a housing for capturing Carpophyllus truncatus beetles.

16. One or more co-attractant compounds; Insecticide; A container for holding captured beetles; and A lid for a container that allows beetles to enter the container. The apparatus according to claim 15, further comprising one or more of the above.

17. below: a. The container for holding the captured beetles is transparent; b. The lid of the container is green. The apparatus according to claim 16, wherein one or both of the above conditions are met.

18. A kit comprising the composition according to any one of claims 1 to 10 or the dispenser and capture device according to any one of claims 11 to 14.

19. One or more co-attractant compounds; and insecticide The kit according to claim 18, further comprising one or more of the above.

20. The co-inducing substance compound is C 1 to C 6 alcohol, C 1 to C 4 aldehyde, indole, and C 1 to C 12 The device according to claim 16 or 17 or the kit according to claim 19, which is selected from esters.

21. The apparatus or kit according to any one of claims 16, 19, or 20, wherein the co-attractant compound is selected from ethanol, isopentyl alcohol, acetaldehyde, isobutanol, 2-methylbutanol, ethyl acetate, isopentyl acetate, isobutyl acetate, 2-phenylethyl acetate, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), methyl benzoate, and (Z)-3-hexenyl acetate.

22. The apparatus or kit according to claim 21, wherein the co-attractant compounds are each in separate containers.

23. The apparatus or kit according to any one of claims 16 and 20 to 22, wherein the co-attractant compound is present in the mixture.

24. The apparatus or kit according to claim 23, wherein the co-attractant mixture comprises ethanol and isopentyl alcohol.

25. The apparatus or kit according to any one of claims 16 to 24, wherein the insecticide is an organophosphate.

26. The apparatus or kit according to claim 25, wherein the insecticide is selected from dichlorvos, thiometon, naled, parathion, malathion, and S-benzyldiisopropylphosphorothiolate (IBP).

27. A method for attracting or capturing Carpophyllus trunkatus beetles, comprising the step of exposing an environment infested with beetles to the composition according to any one of claims 1 to 10, the dispenser according to any one of claims 11 to 14, or the apparatus according to any one of claims 15 to 17 and 20 to 26.

28. A method for monitoring the presence of the beetle Carpophyllus trunkatus, comprising the step of placing a composition according to any one of claims 1 to 10, a dispenser according to any one of claims 11 to 14, or an apparatus according to any one of claims 15 to 17 and 20 to 26 in an environment where the presence of the beetle needs to be monitored.

29. The method according to claim 27, wherein the aforementioned environment where the plague has spread is selected from a nut orchard and a nut stockpile.

30. The method according to claim 28, wherein the environment is selected from a nut orchard, a nut stockpile, and nuts that are ready for export or imported.

31. below: i) The nut orchard is selected from almond orchards, pistachio orchards, walnut orchards, cashew nut orchards, kumiri nut orchards, macadamia nut orchards, and Brazil nut orchards; and / or ii) The nuts in stockpiles, or the nuts ready for export or imported, are selected from almonds, pistachios, walnuts, cashews, kumi nuts, macadamia nuts, and Brazil nuts. The method according to claim 29 or 30, wherein one of the above applies.

32. below: i) The nut orchard is selected from almond orchards, pistachio orchards, and walnut orchards; and / or ii) The nuts are selected from almonds, pistachios, and walnuts. The method according to claim 31, wherein one of the following applies.

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