Improved Biological Formulation for Controlling Brown Marmorated Stink Bugs and Other Harmful Insects.

Abstract

The invention relates to a biological formulation enhanced with bacterial and fungal microorganisms for the control of brown marmorated stink bugs and other harmful insects.

Description

CONTROL OF BROWN STINK BUGS AND OTHER HARMFUL INSECTS BIOLOGICAL FORMULATION DEVELOPED FOR Technical Area The invention is a bacterial and fungal treatment for the control of brown marmorated stink bugs and other harmful insects. It relates to biological formulations developed with microorganisms. State of the Art pests such as Halymorpha halys (Brown Marmorated Stink Bug) and Cimex lectularius (Bed Bug) Various methods have been used in the past in this struggle, but these methods have caused various problems. It has given rise to. Among chemical insecticides, glyphosate-based products and pyrethroids are commonly used. These pesticides were used and aimed to kill harmful insects by targeting their nervous system. However, chemical pesticides contaminate soil, water sources, and air, causing environmental toxicity. This has happened, killing soil microorganisms and disrupting the biological balance of agricultural areas. It has contaminated drinking water sources by mixing with groundwater. It also affects beneficial pollinators like bees. Humans have harmed organisms and led to a decrease in biodiversity in the ecosystem. It causes carcinogenic effects, hormonal disorders, and other health problems on health. As a result, agricultural workers and consumers are exposed to these chemicals and face serious risks. Over time, harmful insects have developed resistance to these pesticides, leading to a situation where the pesticides... It reduced its effectiveness, requiring the use of higher doses. The effect of the drugs was affected by temperature. It has varied depending on environmental factors such as humidity, wind, and rain. In biological control methods, entomopathogenic fungi and bacteria, such as Beauveria, are used. Microorganisms such as Bacillus bassiana and Bacillus thurigensis have been used, but the current biological Since the products usually contain a single entomopathogenic microorganism, different harmful insects It has been insufficient in targeting life stages. Factors such as high temperature or low humidity can affect climate. under certain conditions it has become ineffective, only effective against specific pests, and has a wide range of effects. It has not been able to provide the necessary space. The number of commercially available entomopathogen-based products on the market is quite high. Due to its limited scope, its widespread use has remained restricted. Genetically modified organisms developed through genetic engineering are also harmful. It has been used in pest control, but this approach has led to ethical and legal debates. This has created a lack of trust between consumers and farmers. At the same time, these organisms... The long-term effects on natural ecosystems are uncertain. 1 Physical and mechanical methods involve collecting pests by hand, using traps, or Mechanical methods of destruction have been applied, but these methods require long hours to cover large areas. and required labor, was costly, and was insufficient in controlling the pest population. It remains. In general, the high cost of chemical and biological products is a major concern, especially for small-scale farmers. It has created an economic burden, and chemical methods are not a sustainable solution in the long term. It was unable to provide [the necessary support], and harmful populations rapidly increased when the effect of the pesticide decreased. Furthermore, both... Both chemical and biological methods target not only the intended pests but also other ecosystems. It has also negatively affected organisms. There are various patent and utility model applications related to this subject. For example... US10595533B1 Development of a mixture to combat Cimex lectularius (bed bug). The mixture contains active ingredients such as permethrin, piperonyl butoxide, and glycolic acid. It contains and is sprayed on areas such as beds and upholstery to kill bed bugs and their eggs. It aims to... The article titled “The bed bug problem: Past, present, and future control methods” is Pesticide This is a scientific study published in the journal Biochemistry and Physiology in 2013. The article discusses the control methods used in bed bug control from the past to the present. It analyzes; it examines the effectiveness and limitations of past practices, while considering the present situation. how control strategies are shaped and potential future approaches In short, this study evaluates historical pest control methods (e.g., insecticidal methods). (dust and systematic eradication attempts) are methods commonly used today. advantages and disadvantages, as well as future implementation of integrated and innovative strategies It focuses on its potential. Previous methods of combating harmful insects were hampered by their environmental factors. and has proven insufficient due to biological limitations. Chemical insecticides, when applied In these areas, pollinator bees, beneficial insects, and other ecosystems are found in addition to the target pests. It also caused toxic effects on its components, accumulating in soil and water sources over a long period of time. This has created long-term pollution. These insecticides have carcinogenic and hormonal effects on human health. This has created serious risks such as disorders and exposure for agricultural workers and consumers. This has increased the risk. Furthermore, continuous and intensive use has resulted in an increase in harmful insects. The development of resistance to chemicals leads to decreased effectiveness and the need for progressively higher doses. The effectiveness of chemical methods depends on environmental conditions such as temperature, humidity, wind, and precipitation. 2 This has varied depending on the situation, and this has necessitated large-scale and sustainable control. This has made it difficult to ensure. While biological control agents also have certain advantages, a single entomopathogen... Products containing microorganisms are insufficient in targeting different life stages of pests. they have remained, their effectiveness has rapidly increased in adverse climatic conditions such as high temperature or low humidity. It has lost. Such biological products are generally only effective against a limited number of harmful species. It has been effective, but the limited variety of commercially available products on the market is hindering widespread adoption. This has created an obstacle. Both the environmental and biological negative impacts of chemical insecticides and The limited scope of biological control methods and their susceptibility to climatic conditions make them harmful to pests. It has failed to offer lasting and effective solutions in insect control; harmful insects have developed resistance. The tendency to improve upon existing techniques has led to their failure in the long run. In conclusion, it provides highly effective, environmentally friendly, and climate-controlled solutions for combating harmful insects. resistant to adverse conditions, capable of preventing the development of resistance, and sustainable in the long term. A new technology that offers a solution is needed. Brief Description and Objectives of the Invention The invention aims to offer an environmentally friendly, sustainable, and effective solution for combating harmful insects. It targets, in particular, Halymorpha halys (Brown Marmorated Stink Bug) and Cimex lectularius The shortcomings of current methods used in combating pests such as bed bugs. It was developed with the aim of eliminating and presenting an innovative approach in this field. Chemical By eliminating the harm that insecticides cause to the environment and other living things, biologically... These methods contribute to the protection of natural ecosystems and mitigate toxic effects. It offers a purified solution. The invention targets all life stages of harmful insects, including eggs, larvae, nymphs, and adults. by taking, instead of a single microorganism, Bacillus thurigensis, Beauveria bassiana and Nosema Through the synergistic effect of bacterial, fungal and microsporidial microorganisms such as maddoxi, all It aims to control the life cycle. Developed resistance to chemical insecticides. To eliminate the resistance problem, use a mixture of multiple microorganisms to treat resistance. It reduces the likelihood of further development and offers a long-term solution. Formulated to be effective under different climatic conditions (temperature, humidity, wind, rain, etc.) Thanks to its biologically resistant formulation resistant to environmental factors, it can be used in a wide geographical area. It provides applicability in agricultural areas (orchards, vegetable fields, grain fields). control of harmful insects in (warehouses) and enclosed areas (homes, hotels, hospitals, warehouses) and 3 It makes eradication possible, and also in the fields of environmental protection and public health. It is available for use. By eliminating risks to human health, chemical insecticides can be used in food products and reducing environmental pollution caused by accumulated waste in the environment, and increasing agricultural productivity. By increasing [the number of participants], it offers an economical and efficient solution. With the aim of protecting public health, especially in controlling pests that threaten human health and chemical insecticides It prevents the health problems it causes. Instead of a single microorganism, bacterial, fungal, and microsporidial microorganisms This innovative biological formulation, which utilizes its synergistic effect, is used in the fight against harmful insects. By offering a more effective, safe and environmentally friendly method, it is suitable for both agricultural and indoor use. It provides a wide range of uses and high efficiency in applications. Furthermore, it targets... By exerting a targeted effect, it directly targets various life stages of harmful insects and A lower-cost and more economical solution in the long run compared to chemical insecticides. It provides. The invention eliminates the environmental and biological problems caused by chemical insecticides, It aims to set a new standard in combating harmful insects. Detailed Description of the Invention The invention represents an advancement in the field of biological control and entomopathogenic microorganisms. It is an innovation developed for use in the agriculture, environmental protection, and public health sectors. This relates to formulation, particularly within the scope of biological control against harmful insects. Pests such as Halymorpha halys (Brown Marmorated Stink Bug) and Cimex lectularius (Bed Bug) It aims to control insects. The invention is used in combating harmful insects. an environmentally friendly, sustainable and effective biological formulation and application It includes the method. In this context, especially species such as Halymorpha halys and Cimex lectularius. Environmental and biological problems caused by chemical insecticides in pest control It aims to eliminate bacterial, fungal and microsporidial microorganisms. By utilizing its synergistic effect, it targets all life stages of harmful insects and has a wide range of applications. It offers a range of uses. The essential elements of the invention include bacterial microorganisms, fungal microorganisms, microsporidial microorganisms, carrier substances, special formulations, and application devices. It is located in the section on bacterial microorganisms, specifically Bacillus thurigensis, which is associated with insects. While killing pests by targeting their digestive system, Bacillus subtilis also kills their eggs and 4 It affects the larvae. Bacillus cereus has a high control rate against harmful insects. While targeting nymphs, Bacillus velezensis (B. amyloliquefaciens) It provides effective control by targeting bacteria such as Bacillus brevis, which are harmful insects. It suppresses the immune system, caused by the insect Lysinibacillus sphaericus (Bacillus sphaericus). It targets the larvae and is lethal on the eggs of Pantoea agglomerans beetles. It is having an effect. In the fungal part, Beauveria bassiana attaches to the exoskeleton of insects and infects them. It causes a mortality rate of 76.19%. Lecanicillium lecani affects various life stages of insects. When targeting, Metarhizium anisopliae kills insects by infecting their exoskeleton. In the microsporidial portion, Nosema maddoxi enters the digestive system of harmful insects. They cause infections and reduce populations by disrupting energy metabolism. Carrier materials are used to stabilize microorganisms and maintain their effectiveness. These substances are used to increase the resistance of microorganisms to environmental conditions. It increases efficiency and provides ease of implementation. As part of the technical problem-solving method, the first step is to analyze the life stages of the harmful insects. Bacterial and fungal microorganisms are selected to be targeted. The selected microorganisms... a formulation consisting of two parts (bacterial and fungal) mixed in specific proportions It is prepared and microorganisms are placed in a carrier to form a stable formulation. It is combined with other substances. The formulation consists of two parts: a bacterial mixture. While targeting the symbiotic bacteria and larvae of harmful insects, the fungal mixture targets the pests. They kill insects by infecting their exoskeletons. Microorganisms are present in varying proportions. Mixing them together provides maximum effect. Spray devices are used as application equipment for the formulation to be applied to agricultural areas or indoors. It enables application to various areas; while powder or spray form is suitable for confined spaces and narrow areas. It is applicable. The invention works by effectively targeting all life stages of harmful insects. It provides eradication. Eggs are affected by organisms such as Pantoea agglomerans and Bacillus cereus. Microorganisms are effective, and these microorganisms infect the eggs, affecting their development. It prevents this. Lysinibacillus sphaericus and Beauveria bassiana are effective against the larvae, It kills the larvae by infecting them. The pests in the nymph stage are caused by Bacillus cereus and... It is targeted by Metarhizium anisopliae. In adult insects, it is targeted by Beauveria. Bassiana and Nosema maddoxi are effective by targeting the exoskeleton and digestive system. It provides eradication. Mixtures of different entomopathogenic microorganisms in the preferred applications of the invention. For example, Trichoderma spp. or can be used instead of the microorganisms included in the invention. Other bacterial or fungal species, such as Serratia spp., may be preferred. Another preference of the invention. In its application, it consists of a combination of chemical insecticides and biological agents. Formulations can be used. In addition, harmful insects can be controlled using genetic engineering methods. The use of target microorganisms may also be preferred. In addition, Different carrier substances can be used to increase the stability of microorganisms. For example, nanotechnology-based carriers or polymer-based encapsulation methods are preferred. It can be done. The invention has a wide range of applications in combating harmful insects, both in agriculture and in other fields. effective in enclosed spaces, as well as in environmental, industrial and public health applications. It can be used in the following ways. In agricultural applications, Halymorpha halys (Brown In the fight against pests such as the stink bug, orchards, vegetable fields, grain silos and other It provides effective results in agricultural areas. In this context, biological formulations are used for plants. It can be applied by spraying, given to the soil surface to target larvae. Alternatively, pest control can be achieved in the early stages through seed coating methods. In indoor applications, against household pests such as Cimex lectularius (bed bug). It is used in homes, hotels, hospitals, and warehouses in the fight against the disease. The formulation is used on furniture, beds, carpets, etc. It can be applied to surfaces as a spray, given in powder form to confined spaces, or used for intensely harmful purposes. It can be used in areas where populations are present through fumigation methods. In environmental applications, harmful substances are found in parks, gardens, green spaces, and other open areas. Insect control can be achieved. For this purpose, aerial surveillance can be carried out over large areas using drones or airplanes. It can be applied by spraying or by integrating the formulation into irrigation systems. In industrial applications, it is used to control harmful insects in food storage and processing facilities. Food safety is supported by ensuring this. Spraying can be used in storage areas, as well as in narrow spaces. The powder form can be applied to the areas. In public health practices, mosquitoes, bedbugs, and other pests that threaten human health are addressed. Pests can be controlled. Microorganisms such as Lysinibacillus sphaericus in water Mosquito larvae can be controlled by applying it to puddles, and in enclosed spaces. 6 Pests can be eradicated by spraying. Furthermore, the invention addresses pests that cause agricultural pandemics. It can be used in large-scale area applications for controlling invasive species such as Halymorpha halys. In agricultural pandemics caused by species, biological formulations are widely used through spraying. By applying these methods to the affected areas, effective control measures can be achieved. The invention offers effective eradication by targeting all life stages of harmful insects. With its environmentally friendly design, it stands out as both an economical and sustainable solution. Formulation Examples and Component Ratios (Table 1) The table below shows two different "Examples" of the invention optimized for different application areas. The "Application" prescription has been presented: Bacterial Group: Bacillus thurigensis (targeting the digestive system), Bacillus Bacillus subtilis (targeting eggs / larvae), Bacillus cereus (targeting nymphs), Bacillus velezensis (symbiotic bacteria targeting), Bacillus brevis (immunosuppressive), Lysinibacillus At least 2 selected from between sphaericus (larva targeting) and Pantoea agglomerans (egg targeting). microorganism Fungal Group: Beauveria bassiana (exoskeleton infection and 76.19%). mortality), Lecanicillium lecani and Metarhizium anisopliae (exoskeleton targeted) Microsporidial Group: Nosema maddoxi (energy metabolism disruptor) Table 1: Component Group Example 1: Liquid Formulation (Spray / Pickle) 30% (Equal weight ratios of B. thurigensis, B. subtilis, B. cereus) (Contains bacterial mixture, etc.) Fungal Mixture 30% (B. bassiana, L. lecani, M. anisopliae) Microsporidial Agent 10% (Nosema maddoxi) Carrier Material 20% (Water or Vegetable Oil Based Solvent) Stabilizer / Preservative 10% (Tween 80 or Sodium Alginate) TOTAL 100% Synthesis and Preparation Method The preparation of a biological formulation, which is the subject of this invention, involves maintaining the viability of microorganisms and In order to preserve the synergistic effect, the following specific procedural steps are followed: It is performed: 7 Preparation (Incubation) of Microorganism Cultures  Bacterial Group: Bacillus species and Pantoea agglomerans strains, Nutrient Broth (NB) The culture medium is incubated in an incubator at a constant temperature of 28°C-30°C for 24-48 hours.  Fungal Group: Beauveria bassiana, Lecanicillium lecani and Metarhizium anisopliae The strains were cultured on Potato Dextrose Agar (PDA) or liquid medium at a temperature of 25-27°C for 7-10 days. It is cultured until the sporulation is completed over a period of days.  Microsporidial Group: Nosema maddoxi spores, host cell lines or Purified suspensions are stored at 4C and after the mixing stage It is brought to room temperature just beforehand. Formulation Combining (Mixing Process) The mixing process is "gradual" to prevent microorganisms from experiencing mechanical stress. This is done in the reactor using the "mixing" method:  Step A (Carrier Phase): First, add the main carrier phase (liquid formulation) to the mixing tank. Distilled water is used for this purpose.  Step B (Bacterial Mixture): While the tank is stirred at a constant speed of 100 rpm, the prepared mixture... The bacterial group is added and mixed for 15 minutes to ensure homogeneous distribution.  Step C (Fungal and Microsporidial Mixture): Fungal mixture following bacterial mixture. Sport suspension and Nosema maddoxi are added. Mixing speed damages sport structures. To avoid this, it is kept in the 80-100 rpm range.  Step D (Stabilization): In the final stage, to extend the shelf life of the formulation and protect it from UV rays. stabilizing agents (e.g., 1-2% sodium) to increase resistance Alginate or Tween 80 is added and mixed for another 10 minutes. Environmental Conditions and Control  Temperature Control: Reactor internal temperature throughout the entire mixing process. To prevent microorganisms from experiencing thermal shock, the temperature is kept constant at around 25°C.  Pressure: The process is carried out at normal atmospheric pressure (1 atm).  Quality Control: Samples taken from the final product are tested using the batch dilution method. It is checked and confirmed that each ml / g of product contains at least 10 CFU of live microorganisms. It is done. 8 Application and Field Results The formulation can be applied through integration into spraying or irrigation systems.  Pest Control: Complete eradication of the pest at all stages (from egg to adult). provides.  Plant Growth: Field experiments have shown that this biological mixture has a "fertilizer effect" on the plant. It has been proven to increase product efficiency by 20%. 9

Claims

1. Biological treatment developed for the control of brown marmorated stink bugs and other harmful insects. It is a formulation, and its characteristic is;  Bacterial mixture including Bacillus thurigensis, Bacillus subtilis, Bacillus cereus, Bacillus velezensis (B. amyloliquefaciens), Bacillus brevis, Lysinibacillus At least 2 selected from between Bacillus sphaericus (Bacillus sphaericus) and Pantoea agglomerans microorganism,  Fungal mixture consisting of Beauveria bassiana, Lecanicillium lecani and Metarhizium anisopliae,  Nosema maddoxi as a microsporidial agent,  Water or vegetable oil-based solvent as a carrier substance,  Tween 80 or sodium alginate as a stabilizer includes and In the formulation in question, the bacterial mixture constitutes 30% by weight, and the fungal mixture... 30% by weight of microsporidial agent, 10% by weight of carrier substance, and The stabilizer should be 10% by weight.

2. Biological formulation according to Claim 1, characterized by its bacterial mixture of B. thurigensis, B. It includes Subtilis and B. Cereus.

3. According to Claim 1, it is a biological formulation, characterized by being a liquid or powder formulation. It is a formulation.

4. According to Claim 3, it is a biological formulation, and its characteristic is that the liquid formulation in question... It is in spray form.

5. It is an irrigation system and its characteristic feature is that it contains a biological formulation according to requirement 1.