MXPA97007017A - Novedous pesticide composition and bacillus thuringien seed - Google Patents

Abstract

The present invention relates to a novel strain (s) of Bacillus thuringiensis, in which essentially all the pesticidal activity of said strain is in the supernatant of a fermentation of said strain. The strain produces a substance, which has activity against an insect plague (s) of the order of Coleoptera and which improves the pesticidal activity of a pesticide related to Bacillus. The invention furthermore relates to pesticidal compositions comprising the substance and a pesticide. pesticide vehicle, or the substance and a pesticide related to Bacillus, a chemical pesticide and / or a virus with pesticidal properties, as well as methods for using pesticide compositions for the control of a pesticide.

Description

NOVEDOSE PESTICIDE COMPOSITION AND BACILLUS THURINGIENSIS SEED This application is a continuation in part of the serial request no. 08 / 404,076, filed on March 14, 1995, which is a continuation in part of the serial request no. 08 / 212,462, filed on March 14, 1994, incorporated herein by reference.

FIELD OF THE INVENTION The invention relates to a novel strain (s) of Bacillus thuringiensis, in which essentially all the pesticidal activity is in the supernatant of a fermentation of said strain. The strain produces a substance, which has activity against a plague (s) of insects of the order of Coleoptera and which improves the pesticidal activity of a pesticide related to Bacillus. The invention further relates to pesticidal compositions comprising a substance and a pesticidal vehicle, or the substance and a pesticide related to Bacillus, a chemical pesticide and / or a virus with pesticidal properties, as well as to methods for using the pesticidal compositions to control a plague.

BACKGROUND OF THE INVENTION Each year, significant portions of commercially important agricultural crops in the world, including food, textiles and various domestic plants, are lost by pest infestation, resulting in losses in millions of dollars. Several strategies have been used to try to control these pests. One strategy is the use of broad-spectrum pesticides, that is, chemical pesticides with broad activity. However, there are a number of disadvantages to using such chemical pesticides. Specifically, because of their broad spectrum of activity, these pesticides can destroy non-target organisms, such as insects and beneficial parasites of destructive pests. In addition, these chemical pesticides are often toxic to animals and humans, and target pests often develop resistance when they are repeatedly exposed to these substances. Another strategy has involved the use of biopesticides, which make use of pathogens of natural existence! to control infestations harvested by insects, fungi and weeds. The biopesticides comprise a bacterium, which produces a toxin, a substance toxic to the pest. Biopesticides are generally less harmful to non-target organisms and to the environment as a whole than chemical pesticides. The most widely used biopesticide is Bacillus thuringiensis (B. t.). B. t. It is a spore-forming microorganism, widely distributed, rod-shaped and aerobic. During its sporulation cycle, B. t. produces a protein (s) known as a crystal delta-endotoxin, which annihilates insect larvae. B.t. , therefore, it is very useful as an agricultural pesticide. It has been found that some strains, for example, Bacillus thuringiensis subsp. kurstaki and Bacillus thuringiensis subsp. aizawai, are specific for Lepidoptera. It has been found that Bacillus thuringiensis subsp. israelensis is specific for Diptera (Golgberg, patent of E. U.A. No. 4, 166, 12). It has been found that other strains, for example, Bacillus thuringiensis subsp. tenebrionis (Krieg et al., 1988, U.A. Patent No. 4, 766, 203) is specific to Coleoptera. The isolation of another toxic Bacillus thuringiensis strain of Coleoptera was reported in 1986 (Hernnstadt et al., Bio / Technology vol.4, 305-308, patent of E. U.A. No. 4,764,372, 1988). This strain, designated "Bacillus thuringiensis subsp. San diego", M-7, has been deposited at Northern Regional Research Laboratory, USA under accession number NRRL B-15939. However, the assignee of the '372 patent, Mycogen, Corp., has publicly acknowledged that Bacillus thuringiensis subsp. san diego is Bacillus thuringiensis subsp. tenebrionis. In addition, the '372 patent has been assigned to Novo Nordisk A / S. Additionally, a strain of B. t has been described. , which is toxic against Lepidoptera and Coleoptera (PCT Application No. WO 90/1 3651). The toxin described in the PCT application No. WO 90/13651 has a molecular weight of 81 kd.

During its sporulation cycle, B. t. produces a crystal-like protein (s) known as crystal delta-endotoxin (s), which has a molecular weight of 27-140 kd, which, after ingestion, kills insect larvae. The toxic activity may reside in one or more delta-endotoxins in a strain of B. t. Dadaist. Most delta-endotoxins are protoxins that are proteolytically converted to smaller toxic (truncated) polypeptides in the average intestines of target insects (Hofe and Whiteley, 1989, Microbiol Rev. 53: 242-255). Delta-endotoxins are encoded by cry genes (crystal protein). The cry genes have been divided into six classes and several subclasses based on structural similarities and specific pesticide character. The main classes are, Lepidoptera-specific genes (cryl); Lepidoptera- and Doptera-specific (cryll); Coleoptera-specific (crylll); Diptera-specific (crylV) (Hófe and Whiteley, 1989, Microbiol Rev. 53: 242-255); Coleoptera- and Lepidoptera-specific (referred to as cryV genes by Tailor et al., 1992, Molecular Microbiology 6: 121 1-1217); and Nematode-specific (referred to as cryV and cryVI by Feitelson et al., 1992, Bio / Technology 10: 271-275). Delta-endotoxins have been produced by recombinant DNA methods. The delta-endotoxins produced by recombinant A DN methods may or may not be in the crystal form. The delta-endotoxin of B. t. it is insoluble in water, except at an alkaline pH, and is almost always encoded by a plasmid. It has been shown that some strains of Bacillus thuringiensis produce a heat-stable, pesticide-stable, adenine nucleotide analogue, known as β-exotoxin or thuringiensis, which is only pesticidal (Sebesta et al., In HD Burges (ed.), Microbial). Control of Pests and Plant Diseases, Academic Press, New York, pp. 239-281, 1981). The β-exotoxin has been found in the supernatant of some cultures of Bacillus thuringiensis. It has a molecular weight of 789 and is composed of adenosine, glucose and alric acid (Lüthy et al., In Kurstak (ed.), Microbial and Viral Pesticide, Marcel Dekker, N? Eva York, 1982, pp. 35-72). Its host scale includes, but is not limited to, Musca domestica, Mamestra configurata Walker, Tetranychus urticae, Drosophila melanogaster and Tetranychus cinnabarinus. It is believed that the toxicity of β-exotoxin is due to the inhibition of RNA polymerase directed to DNA through competition with ATP. It has been shown that ß-exotocin is encoded by a plasmid of Cry and five strains of Bacillus thuringiensis K (Bt) and that ß-exotoxin can be classified as ß-exotoxin type I or type II (Levinson et al. 1990, J. Bacterio !. 172: 3172-3179). It was found that ß-exotoxin type I is produced by B.t. subsp. thuringiensis serotype 1, B.t. subsp. tolworthi serotype 9, and B.t. subsp. darmstadiensis serotype 10. It was found that ß-exotoxin type II is produced by B.t. subsp. morrisoni serotype 8ab and is active against Leptinotarsa decemlineata (Colorado potato bugs). Other substances soluble in water that have been isolated from B.t. they include alpha-exotin, which is toxic against the larvae of Musca domestica (Lüthy, 1980, FEMS Microbiol, Lett, 8.1-7); gamma-exotoxins, which are several proteolytic enzymes that include lecithinases, chitinases, and proteases, the toxic effects of which are expressed only in combination with beta-exotoxin or delta-endotoxin (Forsberg et al., 1976, Bacillus thuringiensis: Its Effects on Environmental Quality, National Research Council of Canada, NRC Associate Committee on Scientific Criteria for Environmental Quality, Subcommittees on Pesticides and Related Compounds and Biological Phenomena); lame exotoxin, which has a structure similar to beta-exotoxin, and also active against Leptinotarsa decemlineata (Argauer et al., 1991, J. Entomol. Sci. 26: 206-213); and anhydroturingiensin. { Coll. Czechslovak Chem. Comm. 40, 1775, 1975). WO 94/09630 describes a water soluble substance that improves the activity of Bacillus thuringiensis var. kurstaki and Bacillus thuringiensis var. aizawai Stonard and others (1994, in Natural and Engineered Pest Management Agents, Paul A. Mann, Robert M. Hollingworth, eds. , ACS, Washington, D.C. , p . 25-36) describe diabroticins that have the structure 1 R, RL R2 = H, R3 = OH Diabroticin A 2 R, Ri, R2, R3 = H Diabroticin B Diabroticins were isolated from B. subtilis and have activity against Diabrotica undecimpunctata, Leptinotarsa decemlineata, Anthomus grandis Boheman, mosquito larvae , Staphylococcus aureus, and Micrococcus lutea, but have not had activity against European corn (plague), Escherichia coli, B. subtilis, and Peseudomonas aeruginosa. The activity against other pests was not described by Stonard and others. Diabroticin A was also isolated from the fermentation broths of B. cereus. The technique has strived to obtain an increased mortality of formulations of B.t .. The means have included searching for new strains with increased mortality, strains present by engineering, and designing more effective formulations by combining spores and / or crystals of B.t. with new pesticide vehicles or with chemical pesticides. It is an object of the present invention to improve the insecticidal activity of known formulations of B.t. It is also an object of the present invention to improve the pesticidal activity of pesticides as well as to find novel uses of known pesticidal products. It is advantageous to isolate new strains of Bacillus thuringiensis to produce new substances, so that there is a broader spectrum of biopesticides for any given insect pest.

COMPENDIUM OF THE INVENTION The invention relates to a novel strain of Bacillus thuringiensis in which essentially all the pesticidal activity of said strain is the supernatant of a fermentation of said strain. The crystal protein and spores obtained from a fermentation of a Bacillus thuringiensis strain of the present invention do not possess any pesticidal activity. In a specific embodiment, the strain is selected from the group consisting of EMCC-0077 having the identification characteristics of N RRL B-21090, or mutants thereof having substantially the same properties as EMCC-0077, EMCC-0078 which has the identification characteristics of NRRL B-21090, or mutants thereof having substantially the same properties as EMCC-0078, EMCC-0079 having the identifying characteristics of NRRL B-21092, or mutants thereof having substantially the same properties of EMCC-0079, EMCC-0080 having the identification characteristics of NR RL B-21093, or mutants thereof having substantially the same properties of EMCC-0080, and EMCC-0081 having the identification characteristics of NRRL B -21094, or mutants thereof which have substantially the same properties as EMCC-0081. A substance, which has a pesticidal activity against an insecticide pest of the order of Coleoptera and acts together as, for example, an enhancer or synergist with a pesticide related to Bacillus against a pest is obtained from a supernatant of a fermentation of said strain. In a preferred embodiment, said substance has an LCS0 (LC5o is the concentration of a given pesticidal substance required to kill 50% of the pests) of 126 μg of active ingredient / g of a total material against Leptinotarsa texana. The LC50 of the fermentation pellet of said strain is more than approximately 3000 μg of the active ingredient / g of the total material, as analyzed through bioassay. In another embodiment, said substance has pesticidal activity against an insect pest of the order of Coleoptera. In a more specific embodiment, said substance has pesticidal activity against an insect pest of the species Diabrotica undecimpunctata, Leptinotarsa texana, Anthomus grandia, as well as a surprising activity against a plague of insects of the species Ips calligraphus, Popula japonicus, Epilachna varivastis , Leptinotarsa decemlineata and Dendroctonus frontalis of the order of Coleóptera. In a specific embodiment, said substance improves the insecticidal activity of Bacillus thuringiensis crystal delta-endotoxin (s) against an insect pest (s). In a specific embodiment, said substance improves the insecticidal activity of the crystal delta-endotoxin of Bacillus thuringiensis subsp. tenebrionis against an insect plague (s) of the order of Coleoptera.

As defined herein, "a pesticide related to Bacillus thuringiensis" is a Bacillus strain or spore (e.g., Bacillus thuringiensis or Bacillus subtilis). A pesticide related to Bacillus can also be a substance derived from Bacillus, for example, protein or fragment thereof having activity against or which kills pests; a substance that provides protection to plants, for example, an anti-food substance; or a microorganism capable of expressing a Bacillus gene encoding a Bacillus protein or fragment thereof having activity against or which annihilates pests (eg, Bacillus thuringiensis delta-endotoxin) and an acceptable carrier (see following Compositions section) . The pest may be, for example, an insect, a nematode, a tick, or a snail. A microorganism capable of expressing a Bacillus gene that encodes a Bacillus protein or fragment thereof having activity against or which kills pests that inhabit the phylloplane (the surface of the leaves of plants), and / or the rhizosphere (the soil that surrounds the roots of the plant), and / or aquatic environments, and is able to compete successfully in the particular environment (harvest and other insect habitats) with wild-type microorganisms and provide for the maintenance and stable expression of a gene of Bacillus that encodes a Bacillus protein or fragment thereof that has activity against or that kills pests. Examples of such microorganisms include, but are not limited to, bacteria, for example, the genera Bacillus, Pseudomonas, Erwinia, Serratia, Klebsiella, Xanthomonas, Streptomyces, Rhizobium, Rhodopseudomonas, Methylophilius, Agrobacterium, Acetobacter, Lactobacillus, Arthrobacter, Azotobacter, Lauconostoc , Alcaligenes, and Clostridium; algae, for example, the families of Cyanomophyceae, Prochlorophyceae, Rhodophyceae, Dinophyceae, Chrysphyceae, Prymnesiophyceae, Xanthophyceae, Raphidophyceae, Bacillariophyceae, Eustigmatophyceae, Cryptophyceae, Euglenophyceae, Prasinophyceae, and Chlorophyceae; and fungi, particularly yeast, for example, the genera Sacharomyces, Cryptococcus, Kluveromyces, Sporobolomyces, Rhodotorula, and Aureobasidium. As defined herein, "pesticidal activity" measures the amount of activity against a pest by annihilating or impeding the growth of the pest or protecting the plant from infestation by pests. The invention further relates to pesticidal compositions comprising the substance and a pesticide related to Bacillus, as well as to methods for using the pesticidal compositions to control a pest. The invention is further directed to a method for obtaining a "substantially pure" substance of the present invention comprising the steps of: (a) culturing a Bacillus thuringiensis strain in a suitable growth medium; (b) recovering the supernatant of (a); and (c) subjecting the supernatant from step (b) to column chromatography to purify the substance. As defined herein, a "substantially pure" substance means a substance which contains less than 5% contaminants, for example, the delta-endotoxin protein.

BRIEF DESCRIPTION OF THE DRAWINGS These and other features, aspects and advantages of the present invention will be better understood with respect to the following description, appended claims and appended drawings, in which: Figure 1 shows a synthetic scheme for obtaining structure I. Figure 2 shows the efficacy of the synergism of l a / Ib and NOVODOR ™ on Leptinotarsa decemlineata.

DETAILED DESCRIPTION OF THE INVENTION Obtaining the Substance The substance (s) can be obtained from the supernatant of a Bacillus thuringiensis fermentation including, but not limited to, strains of B. t. of EMCC-0077 having the identification characteristics of N RRL B-21090, or mutants thereof having substantially the same properties as EMCC-0077, EMCC-0078 having the identification characteristics of NRRL B-21090, or mutants thereof having substantially the same properties as EMCC-0078, EMCC-0079 having the identification characteristics of NRRL B-21092, or mutants thereof which have substantially the same properties as EMCC-0079, EMCC-0080 which has the identification characteristics of NRRL B-21093, or mutants thereof which have substantially the same properties as EMCC-0080, and EMCC-0081 which has the characteristics of identification of NRRL B-21094, or mutants thereof having substantially the same properties of EMCC-0081. The substance has activity against an insect pest (s) of the order of Coleoptera and acts together with a pesticide related to Bacillus, such as, for example, an enhancer or synergist. In a specific modality, the substance has the structure (I), wherein, Rt is amino, hydroxy, alkyl alkyl ester (C1.10), aryl ester (for example, benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl), halogen, C1.5 alkoxy, or amino acid, including, but not limited to , alanyl, valinyl, leucinyl, isoleucinyl, phenylalanyl, glycinyl and phenylglycinyl; R2 is amino or alkyl (C? .10); R3 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C? -? O), aryl ester (for example, benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl), halogen, C1.5 alkoxy, methylamine, dimethylamine, thionyl, methyl thionyl, cyano, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R4 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1.10), aryl ester (for example, benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl), halogen, C? S alkoxy, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R5 is hydrogen, methoxy, amino, hydroxy, alkyl alkyl ester (C? -? O), aryl ester (for example, benzoyl, nitrobenzoyl, dinitrobenzoyl, halobenzoyl), halogen, or C? -5 alkoxy; R6 is hydrogen, amino, hydroxy, alkyl (C1.10), ester, halogen or d-s alkoxy; R7 is hydrogen, amino, hydroxy, alkyl alkyl ester (C1.10), aryl ester (for example, benzoyl, nitrobenzoyl, dinitrobenzoyl, halobenzoyl), halogen, C1-s alkoxy, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R8 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1-10), aryl ester (for example, benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl), halogen, C? -S alkoxy, methyl amine, dimethyl amine, thionyl, methyl thionyl, cyano or a salt thereof, including , but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R9 is amino or alkyl (C1.10); and R10 is amino, hydroxy, alkyl (C1.10), alkyl ester (C? .10), aryl ester (for example, benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl), halogen, C1.5 alkoxy, or amino acid including alanyl , valinyl, leucinyl, isoleucinyl, phenylalanyl, glycinyl, and phenylglycinyl. The pyrazine nitrogens can optionally be substituted with alkyl (C1.10), alkyl ester (C? -? 0), aryl ester (for example, benzoyl, nitrobenzoyl, dinitrobenzoyl, halobenzoyl), or oxygen. It should be understood that the invention extends to each of the stereoisomeric forms of the compounds of structure I, as well as to racemates. In highly preferred embodiments, the substance has the structure, hereinafter referred to as "the" or Ib, hereinafter referred to as "Ib".

The: R, RL R2, R3 = H Bacillus thuringiensis can be cultured using means and techniques of fermentation known in the art (see, for example, Rogoff et al., 1969, J. Invertebrate Pah. 14: 122-129; Dulmage and others, 1971, J. Interbrate Path 18: 353-358; Dulmage et al., in Microbial Control of Pests and Plant Diseases, HD Burges, ed., Academic Press, NY, 1980). At the end of the fermentation cycle, the supernatant can be recovered by separating the spores and crystals of B.t. of the fermentation broth by means well known in the art, for example, centrifugation and / or ultrafiltration. Said substance is contained in the supernatant, which can be recovered by means well known in the art, for example, ultrafiltration, evaporation, and spray drying. Alternatively, the substance (s) of the present invention can be obtained through chemical synthesis using methods known in the art. To obtain structure I, the simple pyrazine ring with appropriate substitution and protecting groups can be formed through a number of reactions, for example, spontaneous condensation of alpha-aminocarbonyl compounds. A dihydropyrazine intermediate is formed, but is easily oxidized to pyrazine with oxygen. The dimerization of an individual alpha-aminocarbonyl compound through this method could lead to an individual pyrazine, while a reaction with two different alpha-aminocarbonyl compounds could lead to three products; said substance could be isolated through chromatographic separation (see Figure 1). The latter reaction allows the synthesis of pyrazines with different substituents on each side of the ring. The purification of the substance (s) can be carried out by various methods known in the art, including, but not limited to, chromatography (eg, ion exchange, affinity, size exclusion chromatography / affixing), methods electrophoretic, differential solubility, extraction, or any other normal technique known in the art (see, for example, Protein Purification, eds., JC Janson and Lars Ryden, VCH Publishers, New York, 1989). The activity of said substance can be bioengineered using methods known in the art, such as incorporation of artificial diet, artificial diet coating, leaf paint, leaf drip, and foliar spray. Specific examples of such bioassays are given in the Example section, infra.

Compositions that comprise Substance Said substance can be formulated alone; together with a pesticide related to Bacillus, which, as defined above is a strain, spore, Bacillus protein or fragment thereof that has activity against or that kills pests and optionally an acceptable vehicle in a pesticide composition (s) that is , for example, a suspension, a solution, an emulsion, a dustable powder, a dispersible granule, a wettable powder, an emulsifiable concentrate, an impregnated spray or granule. Examples of said Bacillus strains include, but are not limited to, Bacillus thuringiensis subsp. kurstaki (sold as DIPEL ™ by Abbott Laboratories, Inc., JAVELIN ™ by Sandoz, BIOBIT ™ by Novo Nordisk A / S, FORAY ™ by Novo Nordisk A / S, MVP ™ by Mycogen, BACTOSPEINE ™ by Novo Nordisk A / S, and THURICIDE ™ by Sandoz); Bacillus thuringiensis subsp. aizawai (sold as FLORABAC ™ by Novo Nordisk A / S, and XENTARI ™ by Abbott Laboratories, Inc.); Bacillus thuringiensis subsp. tenebrionis (sold as NOVODOR ™ by Novo Nordisk A / S, TRIDENT ™ by Sandoz, M-TRAK ™ and M-ONE by Mycogen); Bacillus thuringiensis subsp. israelensis (sold either as BACTIMOS ™ or SKEETAL ™ by Novo Nordisk A / S, TEKNAR ™ by Sandoz, and VECTOBAC ™ by Abbott Laboratories, Inc.); Bacillus sphaericus (sold as SPHERIMOS ™ by Novo Nordisk A / S); Bacillus thuringiensis kustaki / tenebrionis (sold as FOIL ™ by Ecogen); Bacillus thuringiensis kurstaki / aizawai (sold as CONDOR ™ by Ecogen and AGREE ™ by Ciba-Geigy); and Bacillus thuringiensis kurstaki / kurstaki (sold as CUTLASS ™ by Ecogen). The Bacillus-related protein can be selected from the group including, but not limited to, Cryl, Cryll, Crylll, CrylV, CryV and CryVI.

Said substance can also be formulated with other factors or substances obtained from the supernatant of a Bacillus supernatant which includes, but is not limited to, an exotoxin and / or the enhancer factor described in the serial application No. 08 / 095,240, filed on 20 July 1993, incorporated herein by reference. Optionally, the formulation may also comprise a pesticide related to Bacillus, chemical pesticide and / or a virus with pesticidal properties and an acceptable vehicle. In a specific embodiment, the components of said composition can act in a synergistic manner. As a result, said composition may have a higher efficiency than can be obtained with each individual component. Synergism can also be manifested by equal or greater efficacy with lower and / or less frequent doses than would be required for each individual component. Alternatively, said substance may act to improve a pesticide related to Bacillus. In compositions comprising the substance and a pesticide related to Bacillus, the substance is present in the amount of about 0.001 to about 300 g per LTU. As defined herein, "LTU" is a unit of Leptinotarsa texana as determined through bioassay. The bioassay compares the sample with a Bacillus reference material, using Leptinotarsa texana or another pest as the normal test organism. Power is determined by dividing the normal reference LC5o and then multiplying by the normal reference power. In another embodiment, the comparison may comprise said substance in a substantially pure form or a supernatant of Bacillus in dry, concentrated or liquid form and a suitable pesticidal vehicle, examples of which are described infra. This composition can be applied separately to a plant, for example, transgenic plants. Specifically, the composition can be applied to a plant previously containing a Bacillus thuringiensis gene. In another embodiment, the composition can be applied to a plant previously exposed to a composition of Bacillus thuringiensis. The substance is present in the composition at a concentration of from about 0.00% to about 60% (w / w). Said compositions described above can be obtained by the addition of a surfactant, an inert carrier, a preservative, a humectant, a food stimulant, a chemical attractant, an encapsulating agent, a binder, an emulsifier, a dye, a UV protector, a pH regulator, a flow agent, or other components to facilitate the handling and application of the product for particular target pests. Suitable surfactants include, but are not limited to, anionic compounds such as a carboxylate, for example, a metal carboxylate of a long-chain fatty acid; an N-acyl sarcosinate; mono- or di-esters of phosphoric acid with fatty alcohol ethoxylates or salts of said esters; fatty alcohol sulfates such as sodium dodecylsulfate, sodium octadecyl sulfate or sodium cetyl sulfate; ethoxylated fatty alcohol sulfates; ethoxylated alkyl phenol sulphates; lignin sulfonates; petroleum sulfonates; alkyl aryl sulfonates such as alkyl benzene sulfonates or lower alkyl naphthalene sulfonates, for example, butyl naphthalene sulfonate; sulfonated salts or condensates of naphthalene formaldehyde; salts of sulfonated phenol-formaldehyde condensates; or more complex sulfonates such as amide sulfonates, for example, the condensation product of oleic acid and N-methyl taurine or the dialkyl sulfosuccinates, for example, the sodium sulfonate or dioctyl succinate. Nonionic agents include, but are not limited to, condensation products of fatty acid esters, fatty alcohols, fatty acid amides or alkyl or alkenyl fatty phenols substituted with ethylene oxide, fatty esters of polyhydric alcohol ethers, for example , sorbitan fatty acid esters, condensation products of said esters with ethylene oxide, for example, polyoxyethylene sorbitol fatty acid esters, block copolymers of ethylene oxide and propylene oxide, acetylenic glycols such as 2,4, 7,9-tetraethyl-5-decin-4,7-diol, or ethoxylated acetylenic glycols. Examples of a cationic surfactant include, for example, an aliphatic mono-, di-, or polyamine such as an acetate, naphthenate or oleate; an oxygen-containing amine such as polyoxyethylene alkylamine amine oxide; amine bonded amine prepared by condensation of a carboxylic acid with a di- or poly-amine; or a quaternary ammonium salt. Examples of inert materials include, but are not limited to, inorganic minerals such as kaolin, mica, gypsum, fertilizer, phyllosilicates, carbonates, sulfates or phosphates; organic materials such as sugar, starches or cyclodextrins; or botanical materials such as wood products, cork, powdered corncobs, rice pods, peanut pods, and ut shells. The compositions of the present invention may be in a form suitable for direct application or as a concentrate or primary composition, which requires dilution with a suitable amount of water or other diluent before application. The pesticide concentration will vary depending on the nature of the particular formulation, specifically, whether it is a concentrate or will be used directly. The composition contains from 1 to 98% by weight of a solid or inert carrier, and from 0 to 50%, preferably from 0.1 to 50% of a surfactant. These compositions will be administered to a regimen marked for the commercial product, preferably from about 0.01 12 to 5.6 kilograms per hectare, when in dry form and from about 0.01 pints to 25 pints per acre, when in liquid form. In a further embodiment, the Bacillus-related pesticide and / or the substance may be treated prior to formulation to prolong the activity of the pesticide when applied to the environment of an objective pest provided that the pre-treatment is not harmful to the pesticide related to the pesticide. Bacillus or substance. Said treatment can be through chemical and / or physical means provided that the treatment does not adversely affect the properties of the composition (s). Examples of chemical reagents include, but are not limited to, halogenating agents; aldehydes such as formaldehyde and glutaraldehyde; anti-infectives, such as zefiran chloride; alcohols, such as isopropanol and ethanol; and histological fixatives, such as Bouin fixative and Helly fixative (see, eg, Humason, Animal Tissue Techniques, W.H. Freeman and Co., 1967). The compositions of the invention can be applied directly to the plant, for example, by spraying or sprinkling at the time when the pest has begun to appear on the plant or before the appearance of pests., as a protection measure. The plants that will be protected within the scope of the present invention include, but are not limited to, cereals (wheat, barley, rye, oats, rice, sorghum and related grains), beets (beet sugar and beet fodder), drupes , handles and soft fruits (apples, pears, plums, peaches, almonds, cherries, strawberries, raspberries and blackberries), leguminous plants (alfalfa, beans, lentils, peas, soybeans), oily plants (turnip, mustard, poppy, olive, sunflowers, coconuts, castor oil plants, cocoa beans, ground nuts), cucumber plants (cucumber, melons), fiber plants (cotton, flax, hemp, jute), citrus fruits (oranges, lemons, grapefruits, tangerines ), vegetables (spinach, lettuce, asparagus, cabbages, and others, carrots, onions, tomatoes, potatoes), lauraceans (avocados, cinnamon, camphor), deciduous trees and with iferos (for example, lime trees, yew trees) , oaks, alders, amos, birch trees, abet We, plants, such as corn, grass plants, tobacco, nuts, coffee, sugar cane, tea, wines, hops, bananas, and natural rubber plants, as well as ornamental plants. The substance can be applied by foliar application, furrow, broadcast grain, "laterally", or soaking the soil. It is generally important to obtain good pest control in the early stages of plant growth, as this is the time when the plant can be severely damaged. The spray or powder may conveniently contain another pesticide, if necessary. In a preferred embodiment, the composition of the invention is applied directly to the plant. The compositions of the present invention can be effective against pests of the order of Coleoptera, for example, Leptinotarsa sp. (for example, Leptinotarsa texana, Leptinotarsa decemlineata), Diabrotica undecimuntata, Dendroctonus frontalis, Anthonomus grandis, Acanthoscelides obtectus, Callosobruchus chinensis, Epilachna varivestis, Pyrrhalta tuteola, Cylas formicarius elegantulus, Listronotus oregonensis, Sitophilus sp. , Cyclocephala borealis, Cycocephala immac ulata, Macrodacrylus subspinosus, Popillia japonica, Rhizotrogu s majalis, Alphitoblus diaperinus, Palorus ratseburgi, Tenebrio m olitor, Tenebrio obscurvus, Tribolium castaneum, Tribolium confusum, Tribolius destructor. The compositions of the invention may also be effective against insect pests including, but not limited to, pests of the order of Lepidoptera, for example, Achroia grisella, Acleris gloverana, Acleris variana, Adoxophyes orana, Agrotis εilon, Alabama argillacea, Alsophilia pometaria, Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis, Archips sp., Argyrotaenia sp. , Atheris mindara, Bombyx mori, Bucculatrix thurberiella, Cadra ca utella, Choridtoneura sp. , Cochylls hospes, Colias eurytheme, Corcyrs cephalonica, Cydia latiferreanus, Cydia pomonella, Datana integerrima, Dendrolimus sibericus, Desmia funeralis, Diaphania hyalinata, Diaphania nitidalis, Diatraea grandiosella, Diatraea saccharalis, Ennomos subsignaria, Eorewma loftini, Ephestia elutella, Erannis tilaria, Estigmene aerial, Eulia salibricola, Eupocoellia ambiguella, Eupoecilia ambiguella, Euproctis chrysorrhoea, Euxoz messoria, Galleria mellonella, Grapholita molesta, American Harrisina, Helicoverpa subflexa, Helicoverpa zea, Heliothis virescens, Hemíleuca oliviae, Homoeosoma eletellum, Hyphantria cunea, Keiferia lycoperscella, Lambdina fiscellaria fiscellana , Lambdina fiscellaria lugubrosa, Leucoma salicis, Lobesia botrana, Loxostege sticticalis, Lymantria dispar, Macalla thyrsisalis, Malocosoma sp. , Mamestra brassicae, Mamestra configurata, Manduca quinquemaculata, Manduca sexta, Maruca testulalis, Elanchra picta, Operopthtera brumata, Orgya sp. , Ostrinia nubilalis, Paleacrita venata, Papilio cresphontes, Pectinophora, Pectinophora gossypiella, Phyrganidia californica, Phyllonorycter blancardella, Pieris rapae, Plathypena scabra, Platynota flouendana, Platynota stultana, Platyptilia carduidactyla, Plodia interpunctella, Plutella xylostella, Pntia protodice, Pseudalatia unip? Ncta, Pseudoplasia includens, Sabulodes aegrotata, Schizura concinna, Sitotroga cerealella , Spilonata acellana, Spodoptera sp. , Thaurnstopoea pityocampa, Tineola bisselliella, Trichoolusia ni, Udea rubigalis, Xylomuges curialis, Yponomeuta padella; Diptera, for example, Aedes sp. , Andes vittatus, Anastrepha ludens, Anastrepha suspensa, Anopheles barben, Anopheles quadrimaculatus, Armigeres subalbatus, Calliphora stygia, Calliphora vicina, Ceratitis capitata, Chironomus tentans, Chrysomya rufifacies, Cochliomyia macellaria, Culex sp. , Culiseta inornata, Dacus oleae, Delia antigua, Delia platura, Delia radicum, Drosophilia melanogaster, Eupeodes corollae, Glossina austeni, Glossina brevipalpis, Glossina fuscipes, Glossina morsitans centralis, Glossina moristans morsitans, Glossina moristans submorsitans, Glossina pallidipes, Glossina palpalis gambiensis, Glossina palpalis palpalis, Glossina tachinoides, Haemagogus equinus, Haematobius irritans, Hypoderma bovis, Hypoderma lineatum, Leucopis ninae, Lucilia cuprina, Lucilia sericata, Lutzomyia longlpaipis, Lutzomyia shannoni, Lycoriella mali, Mayetiola destructor, Musca autumnalis, Musca domestica, Neobellieria so. , Nephrotoma suturalis, Ophyra aenescens, Phaenicia sericata, Phlebotomus sp. , Phormia regina, Sabethes cyaneus, Sarcophaga bullata, Scatophaga stercoraria, Stomaxys calcitrans, Toxorhynchites amoboinensis, Tripteroides bambusa; Acari s, for example, Oligonychus pratensis, Panonycuhs ulmi, Tetranychus urticae; Hymenoptera, for example, Iridomyrmex humilis, Solenopsis invicta; Isoptera, for example, Reticulitermes hesperus, Reticulitermes flavipes, Coptotermes formosanus, Zootermopsis angusticollis, Neotermes connexus, Incisitermes minor, Incisitermes immigrants; Siphonaptera, for example. Ceratophyllus gallinae, Ceratophyllus niger, Nosopsyllis fasciarus, Leptosylla segnis, Ctenocephalides canis, Ctenocephalldes felis, Echicnophaga gallinacea, Pulex irritans, Xenopsylla cheopis, Xenopsylla vexabilis, Tunga penetrans; and Tilenchida, for example, Melodidogyne incognita, Pratylenchus penetrans. The following examples are presented by way of illustration, not by way of limitation.

EXAMPLES MPLO AXIS 1 Cultivation of Various Isolated from B. t.

The subcultures, EMCC-0077, EMCC-0078, EMCC-0079, EMCC-0080, and EMCC-0081, maintained on sloping cultures of Nutrient Broth Agar, were used to inoculate 250 ml of deviated shake flasks containing 50 ml of the medium with the following composition. Liquefied liquor 15 g / L Maltrin- 100 40 g / L Potato starch 30 g / L KH2PO4 1.77 g / L The p H of the medium was adjusted to 7.0 using 10 N NaOH. After inoculation, the stirred flasks were incubated at 30 ° C on a rotary shaker at 250 rpm for 72 hours. The complete culture broths were used to test against Diabrotica undecimpunctata.

EXAMPLE 2 Activity of Diabrotica undecimpunctata in the Complete Cllial Broth of Various Isolates of B. t. 2.5 ml of the complete culture broths, obtained from the previous fermentation, were removed and transferred from the shake flasks to 50 μl polypropylene bioassay tubes. Diabrotica undecimpunctata diet was added to each tube at a fine volume of 25 μl. The diet and the test material were then vigorously mixed and dispensed in bioassay trays to perform the bioassay. Three of six of Diabrotica undecimpunctata were applied on the surface of the "diet". Mylar was embedded on the bioassay trays, and the trays were incubated at 28 ° C without photoperiod. The classification was made in 7 days. Mortality was classified on the seventh day after incubation. SS7 = the size of dead larvae on the seventh day when compared with live larvae, control on the same day as SS7 of 4. SS7 = 3, SS7 = 2, and SS7 = 1 represent the size of the larvae s as 75%, 50% and 25%, respectively, of the live, control larvae of 4. The results are shown below in Table 1. These results indicate that in the 5 strains tested, a mortality of 100% was presented. In addition, the dead larvae were 12.5% of the size of the live control larvae.

TABLE 1 Activity of Diabrotica undecimpunctata in Complete Culture Broths Strain% Mortality SS7 EMCC-0077 100 0.5 EMCC-0078 100 0.5 EMCC-0079 100 0.5 EMCC-0080 100 0.5 EMCC-0081 100 0.5 EXAMPLE 3 Location of Diabrotica undecimpunctata Activity In order to test whether the activity of Diabroticá undecimpunctata is associated with the delta-endotoxin / spores or the supernatant, 2.5 ml of the full culture broths of EMCC-0077, EMCC-0080, EMCC-0081 and NB 125 (a Bacillus thuringiensis subsp. tenebrionis developed under the identical conditions) were centrifuged in a Sorvall RC-5B centrifuge at 15,000 rpm (Sorvall SS34 rotor) for 15 minutes to separate the supernatant and the pellet. The crystal delta-endotoxins plus the spores recovered in the pellet. The delta-endotoxins produced by EMCC-0077 isolated from B. t. Active to Diabrotica undecimpunctata have molecular weights of 66 kD, 29 kD and 12 kD, as determined by SDS-PAG E. The delta-endotoxins produced by E MCC-0078 isolated from B. t. active to Diabrotica undecimpunctata have molecular weights of 1 53 kD, 77 kD, 67 kD. 61 kD, 50 kD, 42 kD, 34 kD, 30 kD, 24 kD, as determined by SDS-PAG E. The delta-endotoxins produced by EMCC-0079 isolated from B. t. Active to Diabrotica undecimpunctata have a molecular weight of 135-145 kD, as determined by SDS-PAG E. The delta-endotoxins produced by EMCC-0080 isolated from B. t. Active to Diabrotica undecimpunctata have molecular weights of 94 kD and 40 kD, as determined by S DS-PAG E. The delta-endotoxins produced by EMCC-0081 isolated from B. t. active to Diabrotica undecimpunctata have molecular weights of 129 kD and 32 kD. as determined by S DS-PAG E. Each supernatant (2.5 ml) obtained from the previous centrifugation is transferred to a 50 μl bioassay tube of polypropylene. The pellet was then resuspended in 2.5 ml sterile distilled water and transferred to a 50 ml bioassay tube of polypropylene. AfterDiabrotica undecimpunctata diet was added to the bioassay tubes, which contained both the supernatant and the resuspended pellet at a final volume of 25 ml. The remaining steps of the bioassay are identical to those described above. The classification is also the same as that described above. The results, presented in Table 2, show that the activity of Diabrotica undecimpinctata of EMCC-0077, EMCC-0080 and EMCC-0081 is present in all supernatants, while the minor activity of Diabrotica undecimpunctata of Bacillus thuringiensis subsp. Known tenebrionis is concentrated in the pellet (spore plus crystal).

TABLE 2 Activity of Diabrotica undecimpunctata in the Supernatant and Pella Strain Fraction% Mortality Points Impediment EMCC-0077 Supernatant 100 0.5 Pella 10 3.0 EMCC-0080 Supernatant 100 0.5 Pella 0 4.0 EMCC-0081 Supernatant 100 0.5 Pella 0 3.0 NB125 Supernatant 0 3.0 Pella 50 1.5 EXAMPLE 4 Activity Against Leptinotarsa texana After filtration through a 0.2μ membrane, the supernatant EMCC-0080, obtained from Example 1, was used to analyze the activity of the beetle. The filtered supernatant was applied to the aubergine foliage at a volume of 187.06 LPH (liters per hectare). The dilutions were 0.1: 1, 1: 4, 1: 8 (supernatant: deionized water: v / v). The larvae of Leptinotarsa texana were exposed to the foliage treated following a normal protocol. Each plant was loaded with 20 larvae of Leptinotarsa texana. The results, presented in Table 3, show that the filtered supernatant of EMCC-0080 is active against Leptinotarsa texana.

TABLE 3 Activity of Leptinotarsa texana in EMCC-0080 Strain Dilution% Mortality EMCC-0080 0 95 1: 1 55 1: 4 20 1: 8 0 Control untreated 0 EXAMPLE 5 Effect of EMCC-0080 v NOVODOR ™ The results shown in Table 3, supra, indicate that the supernatant of EMCC-0080 is not active at a dilution of 1: 8 alone. However, when the foliage of the eggplant was treated with 1.25% or 2.5% of 10X of supernatant of EMCC-0080 concentrate plus 200 μ of NOVODOR ™ (Novo Nordisk A / S, Bagsvaerd, Denmark) per ml. a synergistic effect was obtained as evidenced by the sharp decline of the LC5o and LC90 of NOVODOR ™. The data are presented in Table 4 below.

TABLE 4 Synergistic effect of EMCC-0080 and NOVODOR ™ Sample LCSQ (ua / g) 2 LC_go, (μg / g) 2 Inclination ' NOVODOR ™ 642 4,286 1.55 NOVODOR ™ + 1.25% EMCC-0080 250 1,292 1.79 NOVODOR ™ + 2.5% EMCC-0080 98 490 1.83 1 LCso is defined as the concentration that kills 50% of the target insect population. LC90 is defined as the comicentration that annihilates 90% of the population. The inclination refers to the inclination of a% mortality v. concentration curve log.

EXAMPLE 6 Purification of the Active Substance to Coleoptera Produced Through EMCC-0080 of Cepa B.t.

EMCC-0080 of strain B.t. for 24 hours a ° C in a medium with the following composition in grams per liter at a pH of 7.0: Maltodextrin 40g Soy protein 40g KH2PO4 1.8g K2HPO4 4.5g MgSO4-7H2O 0.3g Metals trace 0.2m Cells and other insolubles were removed of the complete culture broth of EMCC-0080 of strain B. t. The centrifugation was followed by filtration of the resulting supernatant through Celite and a 0.2 μ membrane. The resulting permeate was then concentrated 10 times through evaporation. The purification of the coleopterous active substance (s) of the concentrated 10X permeate was obtained using a four step purification procedure. During the purification, the activity was verified by a surface bioassay of Diabrotica undecimpunctata as described infra, and the purity was determined by capillary electrophoresis as described in Example 8. All chromatographic steps employ detection at 226 nm. Specifically, the surface bioassay was conducted as follows. Concentrated 10X permeate samples were applied to individual wells of a microtiter plate containing 200 μl of solidified artificial insect diet, and then air dried. Two of four neonates of Diabrotica undecimpunctata (corn rootworm, CRW) were placed moderately in each cavity with a brush to paint. The microtiter plates were then sealed with Mylar puncture with nothing holes for air exchange and incubated at 30 ° C and at a humidity of 80%. The classification for the mortality percentage was made in 5 days. In the first step, the concentrated 10X permeate was first purified through column chromatography (cation exchange) of Pharmacia S P Sephadex® C-25 (5 x 30 cm). A sample of 450 ml of concentrated 10X permeate was diluted in 18 liters with deionized water, loaded on the column, which was pre-equilibrated with 20 mM of pH buffer of ammonium acetate at a pH of 5.0. The column was eluted at 18 ml per minute with 5.0 liters of continuous gradient from 20 mM to 0.5 M pH buffer of ammonium acetate at a pH of 5.0. 1 ml fractions were collected, bioassayed, and examined for purity. The active fractions were drained (approximately 150 ml), lyophilized, and re-suspended in deionized water at about 1/5 of the original volume. In the second step, 25 ml of the sample from the first step were loaded onto a size exclusion column BioRad P2 (extra fine) (5 x 100 cm), which was pre-equilibrated with deionized water. The column was eluted at a flow rate of 1 ml per minute with deionized water. The 10 ml fractions were collected, bioanalyzed, and examined for purity through capillary electrophoresis. The active fractions were emptied (approximately 400 ml). In the third step, the 400 ml emptying of the second step was diluted to 16 liters with deionized water. The solution was loaded onto a column (strong cation exchange) of Pharmacia S Sepharose® Fast Flow (5 x 30 cm). which was pre-equilibrated with 20 mM pH regulator of ammonium acetate at a pH of 5.0. The column was eluted at a flow rate of 17 ml per minute with 5.0 liters of continuous gradient from 20 mM to 0.5 M pH buffer of ammonium acetate at a pH of 5.0. The 20 ml fractions were collected, bioanalyzed, and examined for purity. The active fractions were drained (approximately 250 ml) and then lyophilized to dryness to remove the volatile ammonium pH buffer. In the fourth step, the lyophilized emptying of the third step was dissolved in 400 ml of deionized water. The solution was loaded onto a BioRad Chelex® 100 column (0.9 x 30 cm), which was pre-equilibrated with 20 mM of ammonium formate pH regulator at a pH of 4.0. The column was eluted at a flow rate of 5 μl per minute with a step gradient of 2.4 liters of 0.02 - >; 0.1 - 0.2? 0.35? 0.5 - 1.0 μM of p H regulator of ammonium formate at a pH of 4.0. The 20 ml fractions were collected, bioanalyzed, and examined for purity. The active fractions were drained (approximately 300 ml) and then lyophilized to dryness to remove the pH regulator of volatile ammonium formate. Capillary electrophoresis shows that the purified coleoptera active material comprises two compounds, la and Ib.

EXAMPLE 7 Elucidation of Structure of Active Substances to Coleoptera The structures of the compounds la and Ib were elucidated from the spectroscopic data collected in their acetylated derivatives (Derivatives A and B). A mixture of 14 mg of the and Ib was acetylated in 5.0 ml of pyridine with 5.0 ml of acetic anhydride and a crystal of 4-dimethylaminopyridine as a catalyst for 24 hours at room temperature, and then purified by H R PC 18 semi-preparative PLC. A sample of 5 mg in 25 μl was loaded onto the column and eluted at 4 ml per minute with 80% water-20% acetonitrile. The detection is at 254 nm. The NMR spectroscopic data in Derivative A indicate the presence of 14 carbons and 17 protons. However, the mass spectral data suggest a molecular weight of 652 and a formula of C2ßH4oNßOi2 (exact mass, 653.2801, M H +, ca le. 653. 2782). Therefore, it was determined that the compound is symmetric, where only half of the signals were observed through RM N. Several spinning systems were observed through RM N. A substituted central pyrazine ring at positions 2 and 5 is indicated by individual high-field proton bands at 8.6 ppm (H-3 and H-6), which shows long-scale couplings for all carbons in the ring and for the first carbon of the side chain (C-7). The side chain of all three carbon was acetylated in both position 7 and 8 with a methylene in position 9. Carbon was found to have a long-scale correlation for the carbonyl of an ester, and the ester was determined as part of an alanine, which was acetylated in the amino group. The structure of Derivative B differs in a position of Derivative A. In one of the side chains of Derivative B, carbon C-7 no longer was acetylated or bound to oxygen, but was found to be a methylene. The other side chain is identical to that in Derivative A. This difference of only one oxygen was also observed in the mass spectral data. The mass spectral data were obtained with an exact mass of 595.2722 (MH +, cale 595.2727) for Derivative B, indicating a formula of C26H3ßN6 ?? o. The optical densities of Derivatives A and B are as follows: Derivative A [a] D 27 = -6.9 ° C and Derivative B [a] D27 = + 32 ° C. Complete assignments of 1 H and 13 C RM N data were made with ba se in decoupling experiments. COZY, HMQC and H M BC. The assignments are presented in Table 5.

TABLE 5 Data of * H and 13C NMR of Derivative A and Derivative B in D-4 methanol Position? (constants mult. integ. coupling) Derivative? Derivative B? 2 152.9 3 8.6 (S, 1H) 154.7 8.54 (S.1H) 5 144.2 144.4 152.9 6 8.6 (S.lH) 151.1 144.2 7 5.84 (< UHJ * 7.4 Hz) 145.6 2.95 (dilH 74.0 37.5 J = I3. 9.9.2) *, **, f - signals to make intercabiadas The mass spectral data for the mixture of the compounds la and Ib give two ions of 400 and 384. From these data, it was determined that the molecular formula of the compound es CieHíßNßOß and compound Ib is C? 6H2ß 6? 5. The structures of the and Ib are determined by comparing the NMR data of Derivative A and Derivative B with the NMR data of the mixture of the and Ib. The structures of the and Ib are shown below: Ib: R, RL R2, = H. R3 = OH The properties of the compounds la and Ib and their acetylated derivatives are summarized below: Derivative A: Molecular Weight: 652 Empirical formula: C28H4oNßO12 UV (MeOH): 275. 310 nm MS (FAB): (M + H) m / z 653.2801.calc.653 2782 Derivative B: Molecular Weight: 594 Empirical formula: C2ßHjßNßO10 UV (MeOH): 275. 310 nm MS (FAB): (M + H) m / z 595.2722, cale.595.2727 Molecular Weight: 400 Empirical formula: C, ßH2ßNßO6 UV ( MeOH): 275. 310 nm MS (FAB): (M + H) m / z 401 Ib: Molecular Weight: 384 Empirical formula: C, 6H28NßO5 UV (MeOH): 275, 310 nm MS (FAB): (M + H) m / z 385 EXAMPLE 8 Quantification of Compounds la and Ib in Fermentation Broths EMCC-0080 of strain B.t. as described in Example 1. The concentration of the compounds la and Ib in the fermentation broth was determined by capillary electrophoresis. Specifically, a BioRad Biofocus 3000 Capillary Electrophoresis System equipped with an uncoated capillary (50 μm x 50 cm), 0.1 M phosphate at a pH of 2.5, voltage at 20 KV, positive to negative polarity was used for the quantification. detection at 200 nm. The volume of the sample is 30 μl with a second injection of 0.3515 kg / cm2. The analysis time is 10 minutes with the active compounds to the Coleoptera, la and Ib, eluting at 6.0 and 5.9 minutes, respectively. Alternatively, a Beckman P / ACE 2100 Capillary Electrophoresis System equipped with an uncoated capillary (50 μm x 47 cm), 0.1 M pH phosphate buffer at a pH of 2.5, voltage at 20 KV, was used for quantification. polarity from positive to negative, and detection at 200 nm. The volume of the sample is 30 μl with a pressure injection of 10 seconds. The time of the analysis is 10 minutes with the active compounds to Coleoptera, la and Ib, eluting at 7.0 and 6.7 minutes, respectively. The cells and other insolubles were removed from the complete culture broth of EMCC-0080 of strain B. t. m edia nte centrifugation and by filtration through Celite and a membrane of 0.2. The resulting supernatant was analyzed by capillary electrophoresis as described above. The results indicated that the active compounds to Coleoptera, la and Ib, each one is presented at a level of approximately 90 mg per liter of culture broth.

MPLO AXIS 9 Determination of the Power of Compounds la and Ib The relative potency of a crude mixture of the compounds la and Ib (approximately 1: 1 w / w) was determined using Leptinotarsa texana as the test insect and comparing the mortality associated with a normal internal preparation of Bacillus thuringiensis subsp. tenebrionis. Foliar bioassays were performed to determine the potency of a crude mixture of the compounds la and Ib against Leptinotarsa texana. To perform the foliar bioassay, the test and normal materials were weighed in 50 ml centrifuge tubes and suspended with deionized water containing 0.1% Tween®20. 1.200 mg of Bacillus thuringiensis subsp. normal tenebrionis and were suspended to give a final concentration of 12, 000 μg / g. Test samples (ie, NOVO DOR ™ and NOVODOR ™ with compounds la and Ib) were treated in a similar manner unless a speed finding bioassay has shown that the dose delivered is too high or too high to result in a sufficient number of valid data points. If this is the case, the concentration of the primary supply solution was increased or reduced by changing the amount of diluent added to the supply solution. Each sample was then homogenized for 30 seconds using a Virtis Homogenizer, and sound was applied for 20 seconds at 100 Watts using a Braunsonic 1510 ultrasonic homogenizer. Each of these supply solutions was then diluted using a Hamilton Microlab 1000 to give seven dilutions in series consisting of 3,000. 2,000 1333, 857. 545, 364 and 245 μg / ml in a total of 16 ml. Each of these 16 ml solutions was applied to approximately 1858.06 cm2 of eggplant leaves using a Devris Linear Track sprinkler calibrated to deliver 187.06 liters per hectare. The control sheets were sprayed with 16 ml of deionized water. The leaves were air dried and then placed on the rim of a translucent 28.35 g plastic cup containing 5 larvae of Leptinotarsa texana as an instar. Then cardboard lids were placed on the sheet and the lid was compressed in place, cutting a sheet disc and sealing it into the cup. The cups were then inverted and the larvae dripped onto the treated surface of the leaf. Eight cups were prepared for each of the seven serial dilutions. The cups were covered together, were marked, placed on supports, and incubated for 3 days at 30 ° C and at a relative humidity of 65%. These 56 experimental cups and 8 control cups constitute a bioassay. After three days, insect mortality was evaluated. Each glass was given a sharp breath and the larvae that did not move were considered dead. The percentage of mortality was calculated, and the data were analyzed via a parallel probit analysis. LCso, LC90 were evaluated. inclination of the regression lines, coefficient of variation and powers. To determine the potency, a crude mixture of the compounds la and Ib was diluted, bioanalyzed, and compared to a Bacillus thuringiensis subsp. Normal tenebrionis, which has a power of 20,000 LTU / grams. { Leptinotarsa texana Units / gram). The potency results are presented in Table 6, infra, and indicate that the crude mixture is composed of the Ib and (1: 1 p / p) has a power of 75, 555 LTU per g of active ingredient with a LCso of 70 ug per ml (1.8 mg total active ingredient per ml).

TABLE 6 Potency of a mixture of compounds la and Ib Sample LCgj ug / ml Estimated power Mix la / Ib 70 75,555 LTU EXAMPLE 10 Potentiation of Crystal Delta-Endotoxin from Bacillus thuringiensis subsp. tenebrionis The ability of the compounds la and Ib to enhance the insecticidal activity of the crystal delta-endotoxin of Bacillus thuringiensis subsp. tenebrionis against Leptinotarsa texana, adding a crude mixture of the compounds la and Ib to NOVODOR ™ and measuring the LCsoS via parallel probit analysis.

Leaf bioassays were performed against Leptinotarsa texana, as described in Example 9, to determine the level of potentiation gained by adding the compounds la and Ib to NOVODOR ™. A crude mixture of the compounds la and Ib was added to NOVODOR ™. The solutions were serially diluted using the Hamilton Microlab 1000 to provide supply solutions containing NOVODOR ™ at 1000.0, 666.7, 444.4, 2857, 181.8, 121.2 and 80.0 μg / g. two different dilutions of a mixture of the and Ib were prepared resulting in seven dilutions containing 72.0, 48.0, 32.0, 20.6, 13.1. 8.7 and 5.8 μg / g (2.5% v / v with 1000 μg / g of NOVODOR ™); and 36.0, 24.0. 16.0, 10.3. 6.5. 4.4 and 2.9 μg / g (1.25% v / v with 1000 μg / g of NOVODOR ™). Net samples were also prepared (without the compounds la and Ib) and reference substances. The LCsoS of the net samples in pairs were divided between the potential LCso values to give the fold reduction in the LC50 associated with the mixture of the compounds la and Ib. The results are presented in Table 7, infra, and indicate that the crude mixture of the compounds la and Ib activates with potency the insecticidal activity of NOVODOR ™ against Leptinotarsa texana.

TABLE 7 Potentiation of NOVODOR ™ With a Mixture of Compounds la and Ib Contra Leptinotarsa texana Sample LC? P uG / ml Potentiation Factor NOVODOR ™ 642 NOVODOR ™ + 1 .25% la / Ib 250 2.6 NOVODOR ™ + 2.50% la / I b 98 6. 5 EXAMPLE 1 1 Activity of a Mix of Compounds v Ib Contra Escarabai os Ips calliai raph? Sv Dendroctonus frontalis The toxicity of a crude mixture of compounds la and Ib was determined against the beetles Ips calligraphus and Dendroctonus frontalis. 3 ml of a crude solution of the compounds la and Ib (1.8 mg of active ingredient per ml) were added to 5 grams of freeze-dried incense pine phloem and 7 ml of distilled water. A control diet was prepared with 10 ml of water, the diet was divided into 3 petri dishes and 5 to 10 adult impluded adults or newly emerged adult Dendroctonus beetles were placed in each box. Three different batches of the treated diet and the control diet were colonized with 10 to 20 insects. The petri dishes were incubated in the dark at 25 ° C, and the number of dead insects was counted, 4, 7 and 10-12 days after placement in the diet. The data presented are averages of 2 or 3 different replicate studies for each of the insect species. The results for Ips calligraphus are presented in Table 8, infra, and indicate that the crude mixture of compounds l a and Ib is insecticidal.

TABLE 8 Evaluation of a Mix of Compounds la and Ib against Ips calligraphus Treatment Treatment # I Insects # of Deaths% Mortality Post- • Average Avg.

Control 4 20 0 0 Control 7 20 0 0 Control 10 20 1 3 la / Ib 4 20 1 5 la / Ib 7 20 7 35 la / Ib 10 20 20 100 The results for the bioassays of Dendroctonus frontalis are presented in Table 9, infra, and indicate that the mixture of compounds la and Ib is insecticidal.

TABLE 9 Evaluation of a Mixture of Compounds la and Ib against Dendroctonus frontalis Treatment Treatment # Insects # of Mutes% Mortality Post-Days Mean Average Control 4 14-20 0 0 Control 7 14-20 1 7 Control 10-12 14-20 3 16 the / Ib 4 10-20 1 5 the / Ib 7 10-20 1 5 the / Ib 10 20 14 83 AXIS MPLO 1 2 Activity against Popillia japonica (Japanese Bass Scale) The crude mixture of the compounds la and Ib was tested for pesticidal activity against third instar Popillia japonica. Perennial ryegrass roots (11 days old) were immersed in the crude mixture of the compounds la and Ib (1.8 mg of the and Ib per ml) and allowed to dry partially. The larva Popillia japonica was placed a third instar in a pot with several treated roots. After 24 hours, roots and larvae were covered with black Wooster mud soil. The control roots were immersed in water, and the untreated controls consisted of larvae placed directly in the mud on day 1. The boats were incubated in the dark at 25 ° C, and the number of dead larvae was verified after 7, 10, 21, 28, and 36 days, and the corrected control mortality was reported. { (Survival control / survival treatment / Survival control) x 100%} . A total of 25 larvae were used for each treatment. The results, as shown in Table 10, demonstrate that the crude mixture of the compounds la and I b is effective against the third instar Popillia japonica.

TABLE 10 Activity of Compounds la and Ib against Popillia japonica 7 days 10 days 21 days 28 days 36 days Not treated # of death 2 Water control # of death 2 3 8 10 11% control 0 4.3 19 25 30 la and Ib # of death 6 8 13 14 15 26.1 42. 9 45 50 EXAMPLE 13 Activity against Epilachna varivesis (Mexican Bean Beetle) The crude mixture of the compounds la and Ib (1.8 mg per ml) was tested for pesticidal activity against larvae of third-instar Epilachna varivesis. A colony in a cage of Epilachna varivesis, adults, was kept in half-moon beans (bean) from the Burpee shrub in a growth chamber under a photoperiod of 16: 8 at 26.6 ° C and at a relative humidity of 50%. Egg masses were collected and allowed to mature in a petri dish containing a wet cotton wick and half-moon bean leaves. After two days, second instar larvae were collected and used for leaf soaking bioassays. To carry out this bioassay, bean leaves were harvested, and the petiole from a single leaf was pushed through the rubber septum of a florist tube containing 4 ml of water. The sheets were then immersed in serial dilutions on the scale of 0-12% v / v of the crude material containing the compounds la and Ib. Once the leaves dried, 8-10 second instar larvae were placed on each leaf. Insects, larvae and florist tubes were placed in a paper cup of 623.7 g, covered with a fine mesh. The cups were kept in the same growth chamber used to grow the beetle colony. Every two days, the cups were removed from the growth chamber, the larvae were counted, and the larvae were replaced with fresh treated leaves. The tests were finished after 8 days. The results, shown in Table 11, show that the crude mixture of the compounds la and Ib is active against larvae of Epilachna varivesis.

TABLE 1 1 Larval Dose Mortality Response of Epilachna varivesis Days after LC50 95% of Limits Exact Treatment% lower upper 4 5.6 2.58 10.65 6 2. 1 2 3.03 9.37 8 1 .94 0.75 2.81 MPLO AXIS 14 Field Analysis against Leptinotarsa decemfineata (Ch inc he of Colorado Potato) The analysis was conducted against Leptinotarsa decemlineata for control in potatoes (variety Katahdin) with the crude mixture of the compounds la and Ib applied at 50, 100, 150 and 300 grams per acre in combination with NOVODOR ™ applied at 0.5 and 1 .0 quarts per acre, and NOVODOR ™ was also applied only to 0.5, 1.0 and 2.0 quarts per acre. The treatments were applied twice in 7 days with a filling CO2 sprinkler, equipped with 3 hollow cone nozzles of Spray Systems TXVX-12 per row and calibrated to deliver 299.29 liters per hectare at 4.82 kilometers per hour and 3.93 kg / cm2. Each treatment was replicated 4 times in two-row charts (86.36 cm separation) by 7.62 in a random block design. Adult and larvae of Leptinotarsa decemlineata were counted from the above without damaging the foliage over 1 5.24 meters row / graph. The results, as shown in Figure 3, demonstrate that a crude mixture of the compounds I and I b provide significant synergistic activity with N OVODOR ™ in the country. At 0.5 quarts of NOVODOR ™ per acre, a control of 21% was observed, while at 50 grams of the crude mixture of the compounds la and Ib per acre, a control of 13% was obtained. However, when both NOVODOR ™ and the crude mixture of the compounds la and Ib were jointly applied to these regimes, the percentage of control was increased to 81%. Similarly, at 100 grams of the crude mixture of the compounds la and Ib per acre, a control of 28% was obtained, while at 0.5 quarts of NOVODOR ™ a control of 21% was observed, but when both NOVODOR ™ and the crude mixture of the compounds and Ib together with these regimes, the percentage of control was increased to 81%. In addition, when the crude mixture of the compounds I and I b at 50 grams per acre and NOVODOR ™ at 1.0 quarts per acre were applied together, the control percentage was increased to 88%.

DEPOSITS OF ORGANIS MOS The following strains of Bacillus thuringiensis have been deposited in accordance with the Budapest Treaty (Treaty of Budapest) in Agricultural Research Service Patent Culture Collection Northern Regional Research Center (NRRL), 1815 University Street, Peoria, Illinois, 61604, USA. Cepa Access Number Deposit Date EMCC-0077 NRRL B-21090 May 10, 1993 EMCC-0078 NRRL B-21091 May 10, 1993 EMCC-0079 N R R L B-21092 May 10, 1993 EMCC-0080 N R RL B-21093 May 10, 1993 EMCC-0081 N RRL B-21094 May 10, 1993 The strains have been deposited under conditions that ensure that access to the crop will be available during the existence of this patent application to one determined by the Director of Patents and Trademarks to be entitled under 37 CF R. § 1 .14 and 35 U. S. C. § 122. The deposit represents a substantially pure culture of each deposited strain. The deposit is available as required by foreign patent laws in countries where the counterparts of the subject application, or its progeny, are filed. However, it must be understood that the availability of a deposit does not constitute a license to practice the subject invention in derogation of patent rights granted by governmental action. The invention described and claimed herein is not limited in scope by the specific embodiments described herein, since these embodiments claim to be illustrations of various aspects of the invention. Any equivalent embodiments are intended to be within the scope of this invention. In fact, various modifications of the invention, in addition to those shown and described herein, will be apparent to those skilled in the art from the foregoing description. Said modifications are also intended to fall within the scope of the appended claims. Various references are cited herein, the descriptions of which are hereby incorporated by reference in their entirety.

Claims (10)

1 .- A strain of Bacillus thuringiensis, in which essentially all the pesticidal activity of said strain is in the supernatant of a fermentation of said strain.

2. The Bacillus thuringiensis strain according to claim 1, wherein the strain of Bacillus thuringiensis is selected from the group consisting of strain EMCC-0077 having the identification characteristics of N RRL B-21090, or mutants thereof having substantially the same properties of EMCC-0077, strain EMCC-0078 having the identification characteristics of NR RL B-21090, or mutants thereof having substantially the same properties as EMCC-0078, strain EMCC-0079 which has the identification characteristics of NRRL B-21092, or mutants thereof having substantially the same properties of EMCC-0079, strain EMCC-0080 having the identifying characteristics of NR RL B-21093, or mutants thereof having substantially the same properties of EMCC-0080, and strain EMCC-0081 having the identification characteristics of N RR L B-21094. or mutants thereof which have substantially the same properties as EMCC-0081.

3. The strain according to claim 1, wherein a substance having pesticidal activity against an insect pest of the order of Coleoptera and wherein said substance acts together with a pesticide related to different Bacillus against a pest, is obtained from the supernatant of the fermentation of said strain.

4. The strain according to claim 3, wherein the substance has a structure (I), wherein R i is amino, hydroxy, alkyl (C? -? 0), alkyl ester (C 1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl. has halobenzoyl, halogen, C? s alkoxy, or amino acid, including, but not limited to, alanyl, valinyl, leucinyl, isoleucinyl, f in ilala ntlo, glycinyl and phenyl g Meinyl; R2 is amino or alkyl (C i -io); R3 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, C-alkoxy, The methylamine, dimethylamine, thionyl, methyl thionyl, cyano, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R4 is hydrogen, amino, hydroxy. alkyl (C1.10), alkyl ester (C 1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogen-benzoyl, halogen, C? .5 alkoxy, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R5 is hydrogen, methoxy, amino, hydroxy, alkyl (Ci.io), alkyl ester (C1.10), aryl ester. wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dnitrobenzoyl, halogen-benzoyl, halogen, or C1.5 alkoxy; R6 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl. halogenobenzoyl, halogen or C? s alkoxy; R7 is hydrogen, amino, hydroxy, alkyl (Ci-io), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogen-benzoyl), halogen, C.sub.

5.5 alkoxy or a salt thereof, including, but not limited to. phosphate, sulfate, acetate, carbonate and nitrate; RB is hydrogen, amino, hydroxy. alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogen benzoyl, halogen, C? .5 alkoxy, methyl amine, dimethyl amine, thionyl, methyl thionyl, cyano or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R9 is amino or alkyl (C1.10); and R10 is amino, hydroxy. alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenbenzoyl, halogen, C? .5 alkoxy, or amino acid including alanyl , valinyl, leucinyl, isoleucinyl, phenylalanyl, glycinyl, and phenylglycinyl. 5. The strain according to claim 4, wherein R is an amino acid selected from the group consisting of alanyl, valinyl, leucinyl, isoleucinyl, phenylalanyl, glycinyl and phenylglycinyl.

6. The strain according to claim 4, wherein R is an amino acid selected from the group consisting of alanyl, valinyl, leucinyl, isoleucinyl, phenylalanyl, glycinyl and phenylglycinyl.

7. The strain according to claim 4, wherein the substance has the structure, the: R, R i, R 2, R 3 - H Ib: R, R 1 t R 2 = H, R 3 = OH.

8. A method to control an insect pest of a species of the order of Coleoptera selected from the g rup consisting of Leptinotarsa decemlineata, Ips calligraphus, Dendroctonus frontalis, Epilachna varivestis, and Popillia japonica, which comprises exposing the pest to an effective amount for controlling a pest of a pesticidal composition comprising, (a) a substance having pesticidal activity against an insect pest of the order of Coleoptera and acting together with a pesticide related to different Bacillus against a pest, said substance being obtained from a supernatant of a fermentation of a Bacillus strain, in which essentially all the pesticidal activity of said strain is in the supernatant of said fermentation, and (b) a pesticidally effective vehicle.

9. The method of agreement with claim 1, wherein the substance has the structure (I), wherein Ri is amino, hydroxy, alkyl (C? -? 0), alkyl ester (C?.? 0), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogen-benzoyl , halogen, C? s alkoxy, or amino acid, including, but not limited to, alanyl, valinyl, leucinyl, isoleucinyl, phenylalanyl, glycinyl and phenylglycinyl; R2 is amino or alkyl (C i.io); R3 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogen-benzoyl, halogen, alkoxy of C? .s, methylamine, dimethylamine, thionyl, methyl thionyl, cyano, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R is hydrogen, amino, hydroxy, alkyl (C? .10), alkyl ester, aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, ds alkoxy, or a salt of the same, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R5 is hydrogen, methoxy, amino, hydroxy. alkyl (C 1.10), alkyl ester (C 1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogen-benzoyl, halogen, or C1.5 alkoxy; R6 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C 1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen or C1-alkoxy .5; R7 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl), halogen, alkoxy of C1.5, or a salt of the same, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R8 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C 1-10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, alkoxy, C? .5, methyl amine, dimethyl amine, thionyl, methyl thionyl, cyano or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; Rg is amino or alkyl (Ci.io); and R10 is amino, hydroxy, (C1-10) alkyl. alkyl ester (C 1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, C 1-5 alkoxy, or amino acid including alanyl, valinyl, leucinyl, isoleucinyl, phenylalanyl, glycinyl , and phenylglycinyl, and (b) a pesticide related to Bacillus.

10. The method of agreement with claim 1, wherein the substance has the structure the: R. R,, R2, R3 = H Ib: R, R, R2 = H, R3 = OH. 1 1 .- A method to enhance the pesticidal activity of a pesticide related to Bacillus, which comprises exposing the pest to a pesticidal composition comprising, (a) a substance having pesticidal activity against a plague pest of the order of Coleoptera and acting together with a pesticide related to different Bacillus against a pest, said substance being obtained of a supernatant of a fermentation of a Bacillus strain, wherein essentially all the pesticidal activity of said strain is in the supernatant of said fermentation, (b) a pesticidally effective vehicle in an amount sufficient to potentiate the pesticidal activity of said pesticide related to Bacillus. 12. A method to obtain a substantially pure substance, which contains pesticidal activity against an insect pest of the order of Coleoptera and which acts together with a pesticide related to a different Bacillus against a pest, said substance being obtained from a supernatant of a fermentation of a strain of Bacillus thuringiensis in which essentially all the pesticidal activity of said strain is in the supernatant of said fermentation: (a) cultivating a strain of Bacillus thuringiensis, in which essentially all the pesticidal activity of said strain is in the supernatant of a fermentation of said strain in a suitable growth medium; (b) recovering the supernatant of (a); and (c) isolating the substance from the supernatant of step (b) to obtain the substantially pure substance. 13. The method according to claim 1, wherein the substance has the structure (I), wherein Ri is amino, hydroxy, alkyl (Ci.io), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, C1 alkoxy .5, or amino acid, including, but not limited to, alanyl, valinyl, leucinyl, isoleucinyl, phenylalanyl, glycinyl and phenylglycinyl; R2 is amino or alkyl (Ci.io); R3 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C?.? O), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, C? S alkoxy, methylamine, dimethylamine, thionyl, methyl thionyl, cyano, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R4 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C? -? O), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, alkoxy of C1.5, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; Rs is hydrogen, methoxy, amino, hydroxy, alkyl (C 1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, or alkoxy of C?. $; R6 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogen benzoyl, halogen or C? .5 alkoxy; R is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl), halogen, alkoxy of C? .s, or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; R8 is hydrogen, amino, hydroxy, alkyl (C1.10), alkyl ester (C1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogenobenzoyl, halogen, C? S alkoxy, methyl amine, dimethyl amine, thionyl, methyl thionyl, cyano or a salt thereof, including, but not limited to, phosphate, sulfate, acetate, carbonate and nitrate; Rg is amino or alkyl (C1.10); and R10 is amino, hydroxy, alkyl (C1.10). alkyl ester (C 1.10), aryl ester, wherein said aryl is selected from the group consisting of benzoyl, nitrobenzoyl, dinitrobenzoyl, halogen-benzoyl, halogen, C? s alkoxy, or amino acid including alanyl, valinyl, leucinyl, isoleucinyl , phenylalanyl, glycinyl, and phenylglycinyl, and (b) a pesticide related to Bacillus. 14. The method according to claim 1, wherein the substance has the structure Ib: R, R, R2 = H, R3 = OH. SUMMARY The invention relates to a novel strain (s) of Bacillus thuringiensis, in which essentially all the pesticidal activity of said strain is in the supernatant of a fermentation of said strain. The strain produces a substance, which has activity against a plague (s) of insects of the order of Coleoptera and which improves the pesticidal activity of a pesticide related to Bacillus. The invention further relates to pesticidal compositions comprising the substance and a pesticidal vehicle, or the substance and a pesticide related to Bacillus, a chemical pesticide and / or a virus with pesticidal properties, as well as methods for using the pesticidal compositions for the control of a pest.