Capsules for the release of the predator Euborellia annulipes in the biological control of pests.

Description

1 / 9 Capsules for the release of the predator Euborellia annulipes in the biological control of pests. BRIEF DESCRIPTION

[001] The present invention relates to the development of a biodegradable capsule for the release of the predator Euborellia annulipes, aiming at the biological control of insects; thus contributing to environmental preservation, since it degrades naturally over time, minimizing negative impacts on the ecosystem. FIELD OF APPLICATION

[002] The present invention belongs to the field of Agriculture, more specifically to the field of pest control, providing a sustainable way to reduce the use of chemical insecticides, and promoting biological and efficient control through the use of predatory insects in pest management. FUNDAMENTALS OF THE INVENTION

[003] In the current market, there is a scarcity of available methods for releasing predatory insects in the field, mainly limited to the use of parasitoids. Parasitoids are organisms that deposit their eggs or larvae inside other insects, which serve as hosts. Although effective in controlling agricultural pests, the exclusive use of parasitoids may not be the most suitable solution in certain cases, because parasitoids are often specific to the host they attack. This means that if the parasitoid's specific host is not present in the field, it will not be able to perform parasitism, much less control the pest, and alternative control measures may be necessary.

[004] Unlike generalist predators, such as Euborelia annulipes, their presence in the field can offer significant benefits in pest control, as they have the ability to feed on a variety of harmful insect types. However, we face a considerable challenge when it comes to effectively releasing these predators into the environment. This scarcity of methods Petition 870240109894, dated 12 / 24 / 2024, page 7 / 32 The 2 / 9 release efficiency negatively impacts the progress of studies aimed at transforming them into readily marketable biological control agents.

[005] The present invention provides a capsule to allow the safe and effective release of the predator Euborelia annulipes in the field. This capsule offers the capacity for storage, transport and release of predatory insects, ensuring their integrity throughout the process, due to its flattened bottom. It is designed to protect the insects from adverse environmental effects, such as sudden temperature changes, excessive humidity and mechanical damage. In this way, the capsule ensures that the predators arrive at their destination alive and in ideal conditions for release. The invention thus represents a significant step in the advancement of biological control in the field, providing a viable and efficient alternative for the use of predatory insects in pest management. STATE OF THE ART

[006] Document EP3504965B1 refers to a biological pest control device made of biodegradable materials, such as PLA. The capsule in this document comprises two parts: a concave upper part and a container capsule, both of which achieve a firm and secure attachment to the support, for example, a plant branch. The device has holes that allow the passage of odors, substances and / or insects.

[007] Document PH12012000194A1, entitled “EARWIG REARING COMPOSITION, USE THEREOF, METHOD FOR REARING A PREDATORY EARWIG, REARING SYSTEM FOR REARING SAID PREDATORY EARWIG AND METHOD FOR BIOLOGICAL PEST CONTROL ON A CROP” refers to an earwig composition comprising a population of this insect and an artificial diet, which can be used to rear said predatory earwig species or to release predatory earwig species in a crop. The document also discloses methods of biological pest control. Petition 870240109894, dated 12 / 24 / 2024, page 8 / 32 3 / 9 in a culture that makes use of earwig composition, with a composition for raising insects of the order Dermaptera. OBJECTIVES OF THE INVENTION

[008] The present patent application aims to develop capsules that allow the release of the predator Euborellia annulipes, for biological control, with a viable and efficient alternative through the use of predatory insects in pest management. ADVANTAGES OF THE INVENTION

[009] The present invention has the advantage of being made from biodegradable materials. This characteristic ensures that the capsule degrades naturally in the environment, without causing damage to the ecosystem. Its manufacture from reused vegetable fibers also brings additional benefits, as it uses resources that would otherwise be discarded. This sustainable approach not only protects predatory insects but also contributes to reducing environmental impact, promoting a circular economy and the preservation of natural resources. Thus, the capsule represents a fully ecological solution, combining efficiency in biological control with a genuine concern for sustainability. The use of different sources of vegetable fiber in the production of this capsule gives the product remarkable versatility. Different sources of vegetable fibers can be used, such as sugarcane bagasse and coconut fibers.This versatility in available raw materials allows production to be adapted according to regional availability, taking local resources into account. DETAILED DESCRIPTION OF THE FIGURES

[010] Figure 1A shows the side view of poles A and B when joined, where pole A has a diameter (A) of 25 mm and a height (B) of 13 mm. Pole B, with the same diameter (A) of 25 mm, has a height (C) of 10 mm and a flattened base (D) of 12 mm. Figure 1B shows the side view of these poles when separated. Petition 870240109894, dated 12 / 24 / 2024, page 9 / 32 4 / 9

[011] Figure 2A illustrates the external top view of pole A, highlighting its thickness (E) of 5.0 mm, the holes (1) with diameter (G) of 2.0 mm forming a circle (F) with a diameter of 15 mm. Figure 2B depicts the internal top view of pole B, highlighting its thickness (H) of 6.0 mm, and with a depression (2) with an external diameter (I) of 13.0 mm and an internal diameter (J) of 7.0 mm.

[012] Figure 3A shows a longitudinal section (AA) of poles A and B, highlighting in Figure 3B that the depth (L) of the holes (1) is also 2.0 mm, the cavity (K) is 11.0 mm, and the depression (2) has a height (M) of 7.0 mm and a thickness (N) of 3.0 mm for the base of pole B. DETAILED DESCRIPTION OF THE INVENTION

[013] This biodegradable capsule was designed to enable the controlled release of the biological agent Euborelia annulipes, both in field environments and in greenhouses. The underlying purpose is to simplify its introduction and provide it with protection against external environmental influences. The specific characteristics of the capsule were developed to safeguard the integrity of the predator, both during the storage period and at the time of its subsequent release. This approach aims to ensure not only the effectiveness of the predator's introduction, but also its ability to establish itself effectively for biological control purposes.

[014] The underlying reasoning is that the nymphs, upon emerging, could then leave the capsules, dispersing into the surrounding environment. The intention behind this procedure is to maximize the effectiveness of introducing the predator into the ecosystem, thus optimizing efforts in the field of biological control.

[015] For the development of the capsule, a spherical design was adopted, characterized by the presence of a division in its central axis. This division comprises two distinct parts, namely pole A (apex of the capsule) and pole B (base of the capsule).

[016] Pole A has a completely concave shape and stands out due to the presence of nine holes (1), each with a diameter of 2.0 mm and a depth (L) Petition 870240109894, dated 12 / 24 / 2024, page 10 / 32 5 / 9 of 2.0 mm. These holes (1) play a crucial role in oxygenation and in the exit of nymphs after hatching.

[017] Pole B, in turn, has a greater weight and is slightly flattened, which prevents it from rolling when placed on a flat surface. This is especially important, as it prevents the holes located on the top of the sphere from becoming blocked. This feature facilitates handling of the capsule both during and after the placement of the eggs of the predator E. annulipes. Internally, pole B is partially filled and has a depression (2) intended to house the eggs.

[018] The capsules were made by combining six components, additionally incorporating a source of vegetable fiber. Two distinct vegetable fibers were used: coconut and sugarcane. In the production process of these capsules, the following components were used: wheat flour, gypsum, biodegradable white glue, nipagin, water, and soybean oil. It is important to mention that the proportions of these components varied according to the vegetable fiber used, as systematically presented in Table 1.

[019] To obtain the vegetable fibers, the following procedures were carefully carried out: Coconut fiber was derived from the husk of the dried coconut fruit, which was cut and crushed using an electric food processor, resulting in a finely crushed material. In the case of sugarcane, the bagasse was subjected to a drying process for 10 days, followed by the same cutting and crushing process.

[020] After all the cutting and grinding stages, all the fibers were subjected to a sieving process using a 1 mm mesh. This procedure aimed to separate impurities and larger fibers that were not cut, resulting in a suitable and homogeneous material to be used in the manufacture of capsules. Petition 870240109894, dated 12 / 24 / 2024, page 11 / 32 6 / 9

[021] To prepare the mixture for filling the mold in capsule production, proceed as follows: First, combine the plaster with the previously crushed vegetable fiber. Then, moisten this mixture with water and gradually add the glue and wheat flour, aiming to achieve the desired consistency. To give the dough a looser texture, add the oil, finishing the process by incorporating the nipagin.

[022] In the capsule manufacturing process, two distinct types of molds were used: one designed for the creation of pole A and another for pole B. Each mold consisted of four components, consisting of two rigid molds and two flexible molds.

[023] After obtaining a homogeneous mixture, the mass is directed to the molding stage. The procedure begins by inserting the silicone mold inside the rigid mold. This strategy aims to prevent the mass from adhering to the mold, thus simplifying its subsequent removal after the molding process is complete. Consequently, a second flexible silicone mold is placed over the mass, followed by the fitting of the rigid mold. At this point, pressure is applied to ensure that the mass is distributed along the sides of the mold, thus assuming the desired shape.

[024] With this procedure completed, the rigid and flexible molds are removed, and any excess mass residue on the edges is carefully removed. Carefully, the capsule is removed from the rigid mold, detaching the last flexible layer that lined the inside of the rigid mold.

[025] The mold designated for pole A was equipped with nine strategically positioned holes to mark the locations where perforations would be necessary. Using a pointed metal rod two millimeters in diameter, the capsules were perforated and shaped according to the determined patterns. Petition 870240109894, dated 12 / 24 / 2024, p. 12 / 32 7 / 9

[026] On the other hand, the mold intended for pole B stood out for having a flattened base, thus providing the capsule with the shape previously described for specific purposes.

[027] After the molding process, the capsules were subjected to a drying stage in an electric oven at a temperature of 160 °C, maintaining this condition for a period of 15 minutes. Then, the capsules were properly stored in a dry and sanitized environment.

[028] Notably, after the drying procedure was completed, it was observed that the poles of the capsules from different vegetable fibers exhibited variations in their weights. However, a consistent pattern, regardless of the fiber used, pole B was always heavier compared to pole A, as evidenced in Table 2. Examples of embodiments of the invention

[029] This patent application presents remarkable applicability in scenarios where there are currently no methods available for the effective release of the predator Euborelia annulipes. The biodegradable capsule stands out as a solution, enabling the controlled introduction of this biological agent into diverse environments, be they agricultural fields or grow houses. By employing these capsules, the limitations arising from the absence of effective means to release the predator are overcome. In this way, the capsule not only offers an efficient alternative, but also establishes a path for the successful implantation of the predator in the ecosystem, contributing substantially to improving biological control strategies.

[030] The influence of capsules on the hatching rate and emergence of E. annulipes nymphs under controlled temperature (25 ± 1°C) and humidity (70 ± 10%) conditions was investigated. Two types of capsules, derived from coconut fiber and sugarcane, were used, in addition to a control (eggs in Petri dishes). The 10-day-old eggs were distributed into treatments with 10 replicates, consisting of 10 eggs of the predator E. annulipes per replicate. Petition 870240109894, dated 12 / 24 / 2024, p. 13 / 32 8 / 9

[031] Egg masses were placed inside pole B using disinfected instruments, and the capsules were sealed with biodegradable white glue, forming spheres. The capsules, along with the artificial diet and refuge substrate, were packed in plastic containers (22.0x15.0x6.0 cm). The containers were then kept in a BOD incubator at a temperature of 25 ± 2 °C, relative humidity of 70 ± 10%, and a 12-hour photoperiod. The hatching rate of E. annulipes nymphs and the number of nymphs emerging from the capsules were evaluated by counting the number of nymphs that emerged from their capsules. Observations were made every 12 hours for 7 days, after which the capsules were opened and the total hatching rate was evaluated.

[032] The hatching rate of E. annulipes nymphs and the exit from the capsules were evaluated. A hatching rate greater than 90% was observed in all capsules. All nymphs that hatched exited their respective capsules, demonstrating that the capsule dimensions were adequate, since there were no live nymphs retained in the capsules or trapped in the exit holes. The results indicated that the use of capsules for insect release resulted in a gradual exit of the nymphs, with a peak observed 84 hours after the eggs were placed in the capsules.

[033] The timing of nymphal emergence and exit was influenced by variation in egg incubation period and the time nymphs remained together after hatching, feeding on the chorion. The gradual release approach may be useful for optimizing the establishment period of predators in a given environment, taking into account critical developmental stages.

[034] In summary, the capsules used in this experiment proved to be effective in promoting the hatching and emergence of E. annulipes nymphs.

[035] Table 1. Quantification of the components and plant fibers used in the making of a capsule intended for the predator E. annulipes. Vegetable Fiber Coconut Sugarcane Petition 870240109894, dated 12 / 24 / 2024, p. 14 / 32 9 / 9 0.4 g 1.0 g Components Quantities Wheat Flour 1.87g 2.85g Plaster 3.15g 3.5g Biodegradable Glue 2.0g 2.6g Nipagin 1.0g 1.5g Water 3.0 ml 4.0 ml Oil 1.0 ml 1.0 ml

[036] Table 2. Average weight, in grams, of poles A and B of capsules made from coconut and sugar cane fibers. Average weight of capsules Vegetable Fibers Pole A (g) Pole B (g) Coconut 2.60 3.14 Sugarcane 2.57 3.11 Petition 870240109894, dated 12 / 24 / 2024, p. 15 / 32

Claims

1 / 1 CLAIMS 1. CAPSULES FOR RELEASING THE PREDATOR EUBORELLIA ANNULIPES, comprising a spherical body with a division in its central axis, consisting of poles A and B, wherein pole A has nine holes (1) of diameter (G) and depth (L) of 2.0 mm, pole B has a depression (2), characterized by being made by combining wheat flour, plaster, biodegradable glue, Nipagin, water and oil, and additionally incorporating a source of vegetable fiber.

2. Capsules for releasing the predator Euborellia annulipes, according to claim 1, characterized in that the vegetable fiber source is selected from coconut or sugarcane.

3. Capsules for releasing the predator Euborellia annulipes, according to claim 1, characterized by having the base of pole B slightly flattened, in addition to being heavier than pole A, regardless of the plant fiber used.

4. USE OF CAPSULES FOR THE RELEASE OF THE PREDATOR EUBORELLIA ANNULIPES, defined according to claims 1 to 3, characterized by their use in the biological control of pests. Petition 870240109894, dated 12 / 24 / 2024, page 22 / 32

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