Bacillus thuringiensis insecticide formulation

Abstract

The insecticide formulation of the present invention comprises bacillus thuringiensis, polyethylene glycol having a weight average molecular weight of 1,000 to 12,000 g / mol as measured according to gel permeation chromatography, and polyphenols.

Description

Background Art Technical Field

[0002] The present disclosure relates generally to pesticide formulations, and more particularly to pesticide formulations comprising Bacillus thuringiensis. Background Art

[0004] Insecticide formulations for crop defense applications are traditionally sprayed onto crop tissue as part of a crop defense formulation. Traditional insecticide formulations may contain pesticides that are perceived as toxic to humans and can remain on crops after harvest and transfer to the end consumer of such crops. Furthermore, water in the form of rain and irrigation can wash traditional insecticides from crop tissue, contaminating waterways while also leaving crops vulnerable to pests.

[0005] Conventional alternatives to traditional pesticides include bio-based pesticides, which use naturally occurring microorganisms and bacteria to deter and kill pests. One bacterium that has been used in bio-based pesticides is Bacillus thuringiensis. Bacillus thuringiensis is applied to crops in the form of spores and crystalline proteins in pesticide formulations. During the sporulation process of Bacillus thuringiensis, the bacteria produces crystalline proteins that are toxic to certain pests. When insects ingest crop tissues containing Bacillus thuringiensis spores and proteins, the proteins open pores in the insect's digestive tract. The Bacillus thuringiensis spores then pass through the pores, become active in the insect's bloodstream, and reproduce. Rapid bacterial growth in the insect's bloodstream leads to septicemia and insect death.

[0006] Due to its mechanism of operation, Bacillus thuringiensis has many disadvantages when used in crop defense environments. For example, CA2184019A1 details that when Bacillus thuringiensis is exposed to ultraviolet radiation, the effects can be inactivation of crystalline proteins and damage to spore DNA. CN103160449A discloses the use of humic acid to protect Bacillus thuringiensis from the effects of ultraviolet radiation. Furthermore, proteins and spores are easily removed from crop tissues by water in the form of rainwater and irrigation. Since Bacillus thuringiensis relies on the vitality of both proteins and spores for maximum effectiveness, the environments in which Bacillus thuringiensis is used in crop defense applications are challenging.

[0007] Pesticide formulations often contain humectants (e.g., polyethylene glycol), spreads and stickers, rheology modifiers, nutrients, and a variety of other adjuvants that result in complex formulations. Interactions and side reactions often occur between the different adjuvants present in pesticide formulations, which can reduce the efficacy of one or more properties of the pesticide formulation. The use of pesticides such as Bacillus thuringiensis often requires the addition of additional adjuvants with potential side effects to address the known challenges of using pesticides.

[0008] For decades, pharmaceutical science has studied the interactions between phenols and nonionic polymers in formulations. For example, BNKabadi's "Interaction of Nonionic Hydrophobic Polymers with Phenols I" examines the interaction between phenol and polyethylene glycol. Kabadi explains that phenol preferentially interferes with the stabilizing and solubilizing properties of polyethylene glycol by forming hydrogen bonds between the phenol's OH groups and the polyethylene glycol's ether bridges. Hydrogen bonds tend to form hydrophobic macromolecular structures that aggregate and separate benzene and polyethylene glycol. As a result, the individual properties of polyethylene glycol and phenol that each imparts to the formulation are reduced or eliminated.

[0009] Thus, it was surprising to discover insecticide formulations comprising both polyphenols and polyethylene glycol, while also addressing one or more of the traditional shortcomings of Bacillus thuringiensis. Summary of the Invention

[0010] The present invention provides a solution for providing pesticide formulations comprising both polyphenols and polyethylene glycol, while also addressing one or more of the traditional shortcomings of Bacillus thuringiensis.

[0011] The present invention is the result of the discovery that, despite the preferential interaction of polyethylene glycol and phenol demonstrated in the prior art, formulations containing both polyethylene glycol and polyphenols can address the traditional difficulties of using Bacillus thuringiensis in crop defense applications. This discovery is surprising because it is expected that the preferential interaction of polyethylene glycol and polyphenols would result in the aggregation of these components, resulting in reduced or no effect of each of these components on Bacillus thuringiensis. Surprisingly, it has been found that the Bacillus thuringiensis viability of the combined polyethylene glycol, polyphenol, and Bacillus thuringiensis formulations after exposure to rain is approximately the sum of each individual component. Such results are unexpected because the aggregation of polyethylene glycol and polyphenols would be expected to result in Bacillus thuringiensis viability that is less than the sum of the individual components.

[0012] Additionally, it has been found that using a combined polyethylene glycol, polyphenol, and Bacillus thuringiensis formulation, greater than 90% Bacillus thuringiensis viability can be achieved after exposure to UV light using simulated sunlight. Similar to rainfastness, the expected reduction in Bacillus thuringiensis viability due to the aggregation of polyphenols surprisingly did not manifest itself.

[0013] The polyethylene glycol, polyphenol and Bacillus thuringiensis formulations of the present invention are particularly useful as insecticide formulations.

[0014] According to at least one feature of the present disclosure, the pesticide formulation comprises Bacillus thuringiensis, polyethylene glycol having a weight average molecular weight of 1,000 to 12,000 g / mol as measured according to gel permeation chromatography, and a polyphenol. DETAILED DESCRIPTION

[0015] As used herein, the term "and / or," when used in the context of a list of two or more items, means that any one of the listed items may be used alone, or any combination of two or more of the listed items may be used. For example, if a composition is described as comprising components A, B, and / or C, the composition may contain A alone; B alone; C alone; A and B in combination; A and C in combination; B and C in combination; or A, B, and C in combination.

[0016] Unless otherwise indicated, all ranges are inclusive. Subscript values in polymer formulas refer to the molar average of the designated components of the polymer.

[0017] Test methods refer to the most recent test methods as of the priority date of this document, unless the date is expressed as a two-digit hyphenated test method number. Reference to a test method includes reference to both the testing association and the test method number. Test method organizations are referenced by one of the following abbreviations: ASTM refers to ASTM International (formerly the American Society for Testing and Materials); EN refers to European Standard; DIN refers to the German Institute for Standardization; and ISO refers to the International Organization for Standardization.

[0018] As used herein, the term "average molecular weight" is the number average molecular weight and is measured using hydroxyl number analysis as described in ASTM standard D4274.

[0019] As used herein, unless specifically stated to the contrary, "wt %" or "weight percent" or "weight %" of a component is based on the total weight of the composition or article in which the component is included. Unless otherwise indicated, all percentages are by weight.

[0020] Insecticide formulations

[0021] The present invention includes an insecticide formulation comprising Bacillus thuringiensis, polyethylene glycol, and a polyphenol. According to various embodiments, the insecticide formulation consists of water, Bacillus thuringiensis, polyethylene glycol, and a polyphenol. The insecticide formulation can be used in a crop defense formulation, wherein the insecticide formulation is 50% by weight or less of the crop defense formulation.

[0022] polyethylene glycol

[0023] The insecticide formulation includes polyethylene glycol. Polyethylene glycol refers to a compound of the formula H-(O-CH2-CH2) q -OH, wherein q refers to the number of repeating units in the polyethylene glycol polymer. The q value of the polyethylene glycol can be in the range of 20 to 250.

[0024] The weight average molecular weight of the polyethylene glycol can be 1,000 g / mol or greater, or 2,000 g / mol or greater, or 3,000 g / mol or greater, or 3,500 g / mol or greater, or 4,000 g / mol or greater, or 4,500 g / mol or greater, or 5,000 g / mol or greater, or 5,500 g / mol or greater, or 6,000 g / mol or greater, or 6,500 g / mol or greater, or 7,000 g / mol or greater, or 7,500 g / mol or greater, or 8,000 g / mol or greater, or 8,500 g / mol or greater, or 9,000 g / mol or greater, or 9,500 g / mol or greater, or 10,000 g / mol or greater, or 10,500 g / mol or greater, or 11,000 g / mol or greater. g / mol or less, or 4,500 g / mol or less, or 4,000 g / mol or less, or 3,500 g / mol or less, or 3,000 g / mol or less, or 2,000 g / mol or less, as measured by gel permeation chromatography. For example, the weight average molecular weight of the polyethylene glycol can be 3,000 g / mol to 9,000 g / mol, or 4,000 g / mol to 8,000 g / mol, or 5,000 g / mol to 7,000 g / mol, or 6,000 g / mol. Blends of different average molecular weight polyethylene glycols can be used in the pesticide formulation at the same or different weight percentages.

[0025] The polyethylene glycol may be 0.2 wt % to 10 wt % of the insecticide formulation. The insecticide formulation may comprise 0.2 wt % or greater, or 0.5 wt % or greater, or 1.0 wt % or greater, or 1.5 wt % or greater, or 2.0 wt % or greater, or 2.5 wt % or greater, or 3.0 wt % or greater, or 3.5 wt % or greater, or 4.0 wt % or greater, or 4.5 wt % or greater, or 5.0 wt % or greater, or 5.5 wt % or greater, or 6.0 wt % or greater, or 6.5 wt % or greater, or 7.0 wt % or greater, or 7.5 wt % or greater, or 8.0 wt % or greater, or 8.5 wt % or greater, or 9.0 wt % or greater, or 9.5 wt % or greater. %, while at the same time 10 wt % or less, or 9.5 wt % or less, or 9.0 wt % or less, or 8.5 wt % or less, or 8.0 wt % or less, or 7.5 wt % or less, or 7.0 wt % or less, or 6.5 wt % or less, or 6.0 wt % or less, or 5.5 wt % or less, or 5.0 wt % or less, or 4.5 wt % or less, or 4.0 wt % or less, or 3.5 wt % or less, or 3.0 wt % or less, or 2.5 wt % or less, or 2.0 wt % or less, or 1.5 wt % or less, or 1.0 wt % or less, or 0.5 wt % or less of polyethylene glycol.

[0026] Bacillus thuringiensis

[0027] The insecticide formulation comprises Bacillus thuringiensis. As defined herein, "Bacillus thuringiensis" is defined as spores and / or crystalline proteins of the species Bacillus thuringiensis, and includes all Bacillus thuringiensis subspecies that exhibit insecticidal properties. Examples of such subspecies include Kurstaki, Israelensis, and Aizawa. Bacillus thuringiensis can be added to the insecticide formulation as a solid or as part of a liquid formulation. The presence and subspecies of Bacillus thuringiensis are determined by random amplified polymorphic DNA analysis. A commercially available liquid formulation of Bacillus thuringiensis is THURICIDE available from CERTIS USA of Columbia, Maryland. TM Insecticide.

[0028] polyphenols

[0029] The insecticide formulation comprises one or more polyphenols. As used herein, the term "polyphenol" is defined to mean a liquid composed of one or more of humic acid, fulvic acid, and tannic acid. Humic acid, fulvic acid, and tannic acid each include multiple phenolic functional groups, making each a polyphenol. Humic acid is an acidic organic polymer that can be extracted from humic matter in soil, sediment, or aquatic environments. Humic acid is identified by Chemical Abstracts Service (CAS) number 1415-93-6 and has an average chemical formula C 187 H 186 O 89 N9S1. Fulvic acid is a compound with CAS number 479-66-3 and C 14 H 12 Tannic acid is an organic acid with the chemical formula of 1401-55-4 and C 76 H 52 O 46 A blend of humic acid and fulvic acid is used as FLORIS TM Soil nutrients can be obtained commercially from ORGANOCAT, Louisville, Kentucky. Tannic acid can be obtained commercially from SIGMA ALDRICH. The presence of polyphenols in the pesticide formulations was determined by high performance liquid chromatography. The formulation may comprise polyphenols in a concentration of 0.2 wt% or greater, or 0.5 wt% or greater, or 1.0 wt% or greater, or 1.5 wt% or greater, or 2.0 wt% or greater, or 2.5 wt% or greater, or 3.0 wt% or greater, or 3.5 wt% or greater, or 4.0 wt% or greater, or 4.5 wt% or greater, while at the same time being 5.0 wt% or less, or 4.5 wt% or less, or 4.0 wt% or less, or 3.5 wt% or less, or 3.0 wt% or less, or 2.5 wt% or less, or 2.0 wt% or less, or 1.5 wt% or less, or 1.0 wt% or less, or 0.5 wt% or less. The wt% of polyphenols in the pesticide formulation is determined based on the amount of material comprising the polyphenols added to the pesticide formulation and the polyphenol concentration. The polyphenols may include humic acid, fulvic acid, or tannic acid, alone or in any combination to achieve the above-mentioned polyphenol concentrations in the pesticide formulation.

[0030] The polyphenols may have a weight ratio of fulvic acid to humic acid of 2:100 or greater, or 4:100 or greater, or 6:100 or greater, or 8:100 or greater, or 10:100 or greater, or 12:100 or greater, or 14:100 or greater, or 16:100 or greater, or 18:100 or greater, while at the same time being 20:100 or less, or 18:100 or less, or 16:100 or less, or 14:100 or less, or 12:100 or less, or 10:100 or less, or 8:100 or less, or 6:100 or less, or 4:100 or less, or 2:100 or less.

[0031] adjuvant

[0032] The pesticide formulation may include one or more additives or adjuvants. Examples of additives include viscosity modifiers, pH modifiers, herbicides, fungicides, combinations thereof, and the like without departing from the teachings provided herein.

[0033] Crop Defense Formulations

[0034] The insecticide formulation can be used in a crop defense formulation. The crop defense formulation can contain 1 wt% or greater, or 5 wt% or greater, or 10 wt% or greater, or 15 wt% or greater, or 20 wt% or greater, or 25 wt% or greater, or 30 wt% or greater, or 35 wt% or greater, or 40 wt% or greater, or 45 wt% or greater, while at the same time being 50 wt% or less, or 45 wt% or less, or 40 wt% or less, or 35 wt% or less, or 30 wt% or less, or 25 wt% or less, or 20 wt% or less, or 15 wt% or less, or 10 wt% or less of the insecticide formulation. Individual components of the insecticide formulation can be added separately to the crop defense formulation without departing from the teachings provided herein.

[0035] Example

[0036] Material

[0037] The Bacillus thuringiensis formulation used in the following samples was a liquid insecticide comprising 98.35 wt% of a solution of Bacillus thuringiensis subsp. kustarchii ("BT solution"), which was formulated as THURICIDE TM HPC-O biopesticide is commercially available from CERTIS USA, Inc. of Columbia, Maryland.

[0038] The polyphenol used in the following samples was a blend of 5.2 wt% humic acid, 0.5 wt% fulvic acid and the balance water, exemplified as FLORISTM Soil nutrients can be obtained commercially from ORGANOCAT, Inc. of Louisville, Kentucky.

[0039] The PEG used in the following samples is polyethylene glycol having a weight average molecular weight of 6000 g / mol and is commercially available from Sigma-Aldrich.

[0040] Sample preparation

[0041] Comparative Examples ("CE") CE1-CE3 and Inventive Examples ("IE") IE1-IE6 were prepared according to the following procedure.

[0042] CE1 was prepared by neat sampling of the Bacillus thuringiensis formulation.

[0043] CE2 was prepared by combining 2 grams of the Bacillus thuringiensis formulation and 5 wt% PEG based on the weight of CE2 and mixing with a magnetic stir bar.

[0044] CE3 was prepared by combining 2 grams of the Bacillus thuringiensis formulation and 5 wt% of polyphenols based on the weight of CE3 and mixing with a magnetic stir bar.

[0045] IE1 was prepared by preparing a preliminary formulation containing 2.5 wt% PEG, 2.5 wt% polyphenols, and the balance water, based on the weight of the preliminary formulation. The preliminary formulation (1 mL), Bacillus thuringiensis formulation (2 grams), and water (17 grams) were combined and mixed with a magnetic stir bar to provide IE1.

[0046] IE2 was prepared by preparing a preliminary formulation containing 5 wt% PEG, 5 wt% polyphenols, and the balance water, based on the weight of the preliminary formulation. The preliminary formulation (1 mL), Bacillus thuringiensis formulation (2 grams), and water (17 grams) were combined and mixed with a magnetic stir bar to provide IE2.

[0047] IE3 was prepared by preparing a preliminary formulation containing 1.5 wt% PEG and 3.5 wt% polyphenols, and the balance water, based on the weight of the preliminary formulation. The preliminary formulation (1 mL), Bacillus thuringiensis formulation (2 grams), and water (17 grams) were combined and mixed with a magnetic stir bar to provide IE3.

[0048] IE4 was prepared by preparing a preliminary formulation containing 3.5 wt% PEG and 1.5 wt% polyphenols and the balance water, based on the weight of the preliminary formulation. The preliminary formulation (1 mL), Bacillus thuringiensis formulation (2 grams), and water (17 grams) were combined and mixed with a magnetic stir bar to provide IE4.

[0049] IE5 was prepared by preparing a preliminary formulation containing 7.0 wt% PEG and 3.0 wt% polyphenols and the balance water, based on the weight of the preliminary formulation. The preliminary formulation (1 mL), Bacillus thuringiensis formulation (2 grams) and water (17 grams) were combined and mixed with a magnetic stir bar to provide IE5.

[0050] IE6 was prepared by preparing a preliminary formulation containing 10.5 wt% PEG and 4.5 wt% polyphenols, and the balance water, based on the weight of the preliminary formulation. The preliminary formulation (1 mL), Bacillus thuringiensis formulation (2 grams), and water (17 grams) were combined and mixed with a magnetic stir bar to provide IE6.

[0051] Table 1 provides a summary of the weight percentages of the various components of the Comparative Examples and Inventive Examples based on the materials used and sample preparation methods.

[0052] Table 1 :

[0053] BT solution PEG Humic acid Fulvic acid polyphenols water CE1 98.35 0.00 0.00 0.00 0.00 0.00 CE2 79.33 2.02 0.00 0.00 0.00 17.20 CE3 79.21 0.00 2.02 0.24 2.26 17.20 1E1 79.33 1.01 1.01 0.12 1.13 17.20 IE2 66.47 1.69 1.69 0.20 1.89 28.83 IE3 74.25 0.47 1.32 0.16 1.48 22.55 IE4 85.24 1.52 0.65 0.08 0.73 11.09 IE5 75.21 2.68 1.15 0.14 1.29 19.57 IE6 74.21 3.96 1.7 0.2 1.9 18.68

[0054] Test Method

[0055] Rain resistance test

[0056] By cutting 5.08 centimeters ("cm") x 10.16 cm of PARAFILM M TM (From Bemis Company) laboratory film sample and place the sample on a black LENETA TM The rain resistance test was carried out according to the chart (from LENETA COMPANY). TM Wiper (from Kimberly Clark) to wipe PARAFILM M TM Laboratory film samples. Before use in rain fastness testing, CE1-CE3 and IE1-IE6 were diluted to 71 g per liter of water. Using an automated pipette, 15 drops (15 μl to 30 μl) of diluted CE1-CE3 and diluted IE1-IE6 were randomly placed in an array on the corresponding samples, one sample for each IE and CE. The diluted IE and diluted CE were vortexed between each set of 5 drops to maintain composition consistency. The samples were dried in an incubator at approximately 28°C for approximately 1 hour with diluted IE1-IE6 and diluted CE1-CE3.

[0057] Using 2 EXO TERRA TMEXO TERRA MONSOON RS400 RAINFALLSYSTEM with standard nozzle TM Subject each of the dry samples to simulated rain without any expansion. Place the sample 33 cm from the nozzle. Spray water at the sample at a flow rate of 1.5 liters / hour and measure at the sample interface for 5 minutes. Allow the sample to air dry.

[0058] Samples were extracted by cutting each sample so that each of the 15 dry droplets representing one IE or CE per sample was centered on approximately 0.63 square centimeters. The fifteen 0.63 square centimeters obtained for each sample were placed in glass vials. Sodium lauryl sulfate solution (1 ml, 2 wt% sodium lauryl sulfate in water) was added to each vial. Each vial was sonicated three times and allowed to soak for 8 hours.

[0059] The residual protein concentration was determined by bicinchoninic acid (BCA) as follows. TM BCA Protein Assay Reagent A and PIERCE TM BCA Protein Assay Reagent B (both obtained from Thermo Scientific) was a combination of Reagent A (2 ml) and Reagent B (40 μl) to form a reagent mixture. One hundred (100) μl of each extracted sample was placed in a corresponding cuvette; the reagent mixture (2 ml) was then added to each cuvette; and the cuvettes were then incubated at 30°C for approximately 2 hours. The CARY 100 from AGILENT was used. TM The residual protein concentration was determined by measuring the absorbance at 562 nm using a UV-visible spectrophotometer.

[0060] By using 1wt% TWEEN TM 20% polysorbate nonionic surfactant solution was used to extract samples from the samples to determine spore viability. TM The samples were diluted with 20% polysorbate nonionic surfactant solution, and then the extracted CE1 and IE1 samples were serially diluted at appropriate concentrations and plated. The extracted and diluted CE1 and IE1 samples were evenly plated in 10 μL drops on agar sample growth plates. The plates were incubated at 30°C for 12 hours. The number of colonies, expressed as log colony-forming units / mL, was counted, taking into account the dilution factor.

[0061] Ultraviolet ("UV") Testing

[0062] Bacillus thuringiensis activity was determined before and after exposure of CE1 and IE1 to light as follows.

[0063] 30 μL of CE1 and IE1 droplets were placed on separate plastic Petri dishes using an automatic pipette and allowed to dry for approximately 1 hour. TM HB175 lamp, CE1 and IE1 were exposed to 35 mW / cm2 light for 2 hours. 1 wt% TWEEN TM CE1 and IE1 were extracted from the culture dish using 20% polysorbate nonionic surfactant solution. TM The samples were diluted with 20 μL of polysorbate nonionic surfactant solution, and then serially diluted at appropriate concentrations and plated. The extracted and diluted CE1 and IE1 samples were evenly plated in 10 μL drops on agar sample growth plates. The plates were incubated at 30°C for 12 hours. The number of colonies, expressed as log colony-forming units / mL, was counted, taking into account the dilution factor.

[0064] result

[0065] Table 2 provides the protein retention of CE1-CE3 and IE1-IE6 at given exposure times under simulated rainfall water conditions.

[0066] Table 2 :

[0067] Exposure time (minutes) Protein retention rate (%) CE1 5 0.00 CE1 10 0.00 CE2 5 61.00 CE3 5 33.00 IE1 5 73.80 IE1 10 65.25 IE2 5 88.89 IE2 10 64.89 IE3 5 93.20 IE3 10 88.14 IE4 5 80.79 IE4 10 56.34 IE5 5 85.08 IE5 10 54.43 IE6 5 92.53 IE6 10 62.53

[0068] As shown in the results, CE1, which represents only the application of Bacillus thuringiensis to crops, exhibited zero protein retention regardless of exposure time, indicating that the Bacillus thuringiensis crystalline protein has little rainfastness. CE2 and CE3 demonstrate that adding polyethylene glycol and polyphenols individually to the Bacillus thuringiensis formulations increases the rainfastness of the crystalline protein. As described above, conventional understanding of phenol and nonionic polymer systems suggests that the phenol and nonionic polymers aggregate via hydrogen bonding, resulting in a decrease in the dispersion of the two components throughout the system. Rainfastness is expected to be correspondingly less than the cumulative addition of the two components due to the expected aggregation and separation. As surprisingly discovered, the rainfastness of IE1-IE6 demonstrates the cumulative properties of the polyphenols and polyethylene glycol. Therefore, IE1-IE6 demonstrate that formulations of Bacillus thuringiensis, polyethylene glycol, and polyphenols can exhibit effective rainfastness by retaining a higher percentage of crystalline protein than any of CE1-CE3.

[0069] Table 3 provides the viability of Bacillus thuringiensis spores after exposure to simulated rainfall water conditions over a period of time.

[0070] Table 3 :

[0071]

[0072] As shown in the results, after rainfastness testing, CE1, which represents the application of Bacillus thuringiensis to crops alone, exhibited a Bacillus thuringiensis spore viability of 68%. IE1 demonstrates that adding both polyethylene glycol and polyphenols to Bacillus thuringiensis surprisingly increased the viability of Bacillus thuringiensis spores after exposure to water. This result demonstrates that the combined polyphenol and polyethylene glycol system not only effectively increased the rainfastness of the crystalline protein, but also effectively maintained the viability of Bacillus thuringiensis spores.

[0073] Table 4 provides the viability of Bacillus thuringiensis spores after exposure to simulated light reduction conditions for a period of time.

[0074] Table 4 :

[0075] Bt activity (log CFU / mL) - dark Bt activity (log CFU / mL)-exposure vitality% CE1 8.53±0.13 5.78±0.10 0 IE1 8.28±0.11 8.04±0.13 94

[0076] As shown in the results, after exposure to simulated sunlight, CE1, which only represents the application of Bacillus thuringiensis to crops, exhibited 0% spore viability. As mentioned above, the traditional understanding of phenol and nonionic polymer systems suggests that phenol and nonionic polymers will aggregate through hydrogen bonding, resulting in a decrease in the dispersion of the two components in the entire system. It is expected that the UV protection provided by the polyphenols will be minimized or eliminated due to aggregation and separation with polyethylene glycol. As surprisingly found, the viability of IE1-IE6 indicates that the polyphenols are still actively protecting Bacillus thuringiensis. Therefore, IE1-IE6 demonstrate that the formulation of Bacillus thuringiensis and polyethylene glycol can also exhibit effective UV protection.

Claims

1. A pesticide formulation comprising: Bacillus thuringiensis; 0.2 wt% to 10 wt% of polyethylene glycol, based on the total weight of the pesticide formulation, wherein the polyethylene glycol has a weight average molecular weight of 1,000 to 12,000 g / mol as measured by gel permeation chromatography; and 0.2 wt% to 5.0 wt% of polyphenols based on the total weight of the pesticide formulation, wherein the polyphenols include fulvic acid and humic acid, wherein the weight ratio between fulvic acid and humic acid is 2:100 to 20:100, and wherein the polyphenols do not include tannic acid.

2. The pesticide formulation according to claim 1, wherein the polyethylene glycol has a weight average molecular weight of 5,000 to 7,000 g / mol as measured by gel permeation chromatography.

3. The pesticide formulation according to claim 2, wherein the polyethylene glycol has a weight average molecular weight of 6,000 g / mol as measured by gel permeation chromatography.

4. The pesticide formulation according to any one of claims 1 to 3, wherein the pesticide formulation comprises 0.5 wt% to 2.0 wt% of polyphenols based on the total weight of the pesticide formulation.

5. The pesticide formulation according to any one of claims 1 to 3, wherein the pesticide formulation comprises 0.2 wt% to 5 wt% of polyethylene glycol based on the total weight of the pesticide formulation.

6. The pesticide formulation according to any one of claims 1 to 3, wherein the Bacillus thuringiensis comprises Bacillus thuringiensis subsp. kustakii.

7. The pesticide formulation according to any one of claims 1 to 3, wherein the weight ratio between fulvic acid and humic acid is 8:100 to 14:100.

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