Macrolide compounds

Abstract: Fermentation product A83543 (1), comprising major components A83543A and A83543D and minor components A83543B, A83543C, A83543E, A983543F, A83543G, A83543H and A83543J, is produced by a newly described species, Saccharopolyspora spinosa . The A83543 components and their acid-addition salts (A83543 compounds) are useful as insecticides, particularly against Lepidoptera and Diptera species. Insecticidal, miticidal or ecto­ parasiticidal combinations, compositions and methods are provided. wherein R is H or a group selected from R² is R¹, R³, R⁵ and R⁶ are hydrogen or methyl; R₄ is methyl or ethyl; or an acid addition salt thereof the compounds when R is other than hydrogen.

Description: This invention relates to new group of macrolide compounds that have insecticidal and miticidal activity. There is a serious need for new insecticides and miticides because target organisms are rapidly developing resistance to insecticides and miticides in current use. Resistance to insecticides in arthropods is widespread, with at least 400 species being resistant to one or more insecticides. Development of resistance to older insecticides, such as DDT, the carbamates, and the organophosphates, is well known. Resistance has even developed, however, to some of the newer pyrethroid insecticides and miticides. A need exists, therefore, for new insecticides and miticides. The control of ectoparasites, such as fleas, ticks, biting flies and the like, has long been recognized as an important problem in animal husbandry. The traditional treatments for domestic animals were topically applied insecticides, such as the famous dips for sheep. Indeed, such treatments are still in wide use. Currently, however, the thrust of research has been to find compounds which can be administered to the animals, especially orally, and which will control ectoparasites by poisoning the parasite when it ingests the blood of the treated animal. This invention relates to a new fermentation product designated "A83543", which is comprised of individual components A83543A, A83543B, A83543C, A83543D, A83543E, A83543F, A83543G, A83543H and A83543J. A83543 and the individual A83543 components are useful for the control of insects, particularly Lepidoptera species, such as Southern armyworm, and Diptera species, such as blow fly, stable fly and mosquito, thus, providing useful insecticidal compositions and methods for reducing population of insects or mites using A83543 compounds. The A83543 compounds of this invention are compounds of formula 1 : wherein R is H or a group selected from R² is R¹, R³, R⁵ and R⁶ are hydrogen or methyl; and R₄ is methyl or ethyl; or an acid addition salt thereof the compounds when R is other than hydrogen. The preferred compounds of the invention, from the point of view of ease of preparation, are those wherein R, R¹, R³, R⁴, R⁵ and R⁶ are present in one of the following combinations: R R₁ R³ R⁴ R⁵ R⁶ (a) Me H Et Me Me (b) Me H Et Me Me (c) Me H Et Me Me (a) Me Me Et Me Me (a) Me H Me Me Me (a) H H Et Me Me (d) Me H Et Me Me (a) Me H Et H Me (a) Me H Et Me H H Me H Et Me Me H Me Me Et Me Me H Me H Me Me Me H H H Et Me Me H Me H Et H Me H Me H Et Me H or an acid addition salt thereof the compounds when R is other than hydrogen. The aminosugar in A83543A has been shown to be β-D-forosamine; and the neutral sugar in A83543A is α-2,3,4-tri-O-methylrhamnose. Nine A83543 components have been characterized. These components are the formula 1 compounds wherein R is other than H. They have the following structures: Component R R¹ R³ R⁴ R⁵ R⁶ A (a) Me H Et Me Me B (b) Me H Et Me Me C (c) Me H Et Me Me D (a) Me Me Et Me Me E (a) Me H Me Me Me F (a) H H Et Me Me G (d) Me H Et Me Me H (a) Me H Et H Me J (a) Me H Et Me H The amino sugar can be removed from the A83543 components to give pseudoaglycones (compounds of formula 1 where R = H). Components A, B, C and G have a common pseudoaglycone (the A83543A pseudoaglycone or pseudo-A). A83543A pseudoaglycone was later found to be produced naturally as an A83543 component. Components D, E, F, H and J each have a unique pseudoaglycone. The A83543 pseudoaglycones have the following structures: Pseudoaglycone R¹ R³ R⁴ R⁵ R⁶ A83543A Me H Et Me Me A83543D Me Me Et Me Me A83543E Me H Me Me Me A83543F H H Et Me Me A83543H Me H Et H Me A83543J Me H Et Me H The pseudoaglycones are useful as intermediates, for example, to the A83543 components. The following paragraphs summarize the physical and spectral properties of the A83543 components and pseudoaglycones. In the discussions infra , the following abbreviations are used: EI-MS: electron-impact mass spectrometry FAB-MS: fast-atom-bombardment mass spectrometry FD-MS: field-desorption mass spectrometry HPLC: high performance-liquid chromatography IR: infrared NMR: nuclear magnetic resonance UV: ultraviolet Characteristics of A83543A Molecular Weight: 731 Empirical Formula: C₄₁H₆₅NO₁₀ FD-MS: see Fig. 4 FAB-MS(M+1): Found: 732.4706; Calcd. C₄₁H₆₆NO₁₀ = 732.4687 (see fig. 8) EI-MS: Found: 731.4612; Calcd. 731.4608 (see fig. 10) UV (EtOH) λmax: 243 nm (ε 8,920), IR (CHCl₃): ν (lactone) 1713; (conjugated ketone) 1657; multiple peaks for C-H vibrations around 2940 and for C-O vibrations around 1060 cm⁻¹ (see fig. 1) -121.8° ( c 1.03, CHCl₃) +6.8° ( c 1.03, CHCl₃) Table I summarizes the ¹H and ¹³C NMR data observed with A83543A (in acetone-d₆). Characteristics of A83543B Molecular Weight: 717 Empirical Formula: C₄₀H₆₃NO₁₀ FAB-MS: (see fig. 9) Characteristics of A83543C Molecular Weight: 703 Empirical Formula: C₃₉H₆₁NO₁₀ FD-MS: (see fig. 5) Characteristics of A83543D Molecular Weight: 745 Empirical Formula: C₄₂H₆₇NO₁₀ UV (EtOH) λmax: 244 nm (ε 9,910) IR (CHCl₃): ν (lactone) 1708; (conjugated ketone) 1658; multiple peaks for C-H vibrations around 2940 and for C-O vibrations around 1070 cm⁻¹ (see fig. 2) -142.9° ( c 1.02, CHCl₃) -29.9° ( c 1.02, CHCl₃) FD-MS: see Fig. 6 Table II summarizes the ¹H and ¹³C NMR data observed with A83543D (in acetone-d₆). Characteristics of A83543E Molecular Weight: 717 Empirical Formula: C₄₀H₆₃NO₁₀ FAB-MS (M+1): Found: 718.4526; Calcd. C₄₀H₆₄NO₁₀ = 718.4530 UV (EtOH) λmax: 244 nm (ε 8,600) IR (KBr) (see fig. 13) Table III summarizes the ¹H and ¹³C NMR data observed with A83543E (in acetone-d₆). Characteristics of A83543F Molecular Weight: 717 Empirical Formula: C₄₀H₆₃NO₁₀ FAB-MS (M+1): Found: 718.4534; Calcd. C₄₀H₆₄NO₁₀ = 718.4530 UV (EtOH) λmax: 243 nm (ε 10,500) and 282 nm (ε 109) IR (KBr): (see fig. 14) Table IV summarizes the ¹H and ¹³C NMR data observed with A83543F (in acetone-d₆). Characteristics of A83543G Molecular Weight: 731 Empirical Formula: C₄₁H₆₅NO₁₀ FAB-MS (M+1): Found: 732.4661; Calcd. C₄₁H₆₆NO₁₀ = 732.4687 UV (EtOH) λmax: 243 nm (ε 8,970) IR (KBr): (see fig. 15) Table V summarizes the ¹H and ¹³C NMR data observed with A83543G (in acetone-d₆). Characteristics of A83543H Molecular Weight: 717 Empirical Formula: C₄₀H₆₃NO₁₀ UV (EtOH) λmax: 243 nm (ε 10,100)* IR (KBr): see fig. 16 Characteristics of A83543J Molecular Weight: 717 Empirical Formula: C₄₀H₆₃NO₁₀ UV (EtOH) λmax: 243 nm (ε 10,100)* IR (CHCl₃): see fig. 16 A83543H and J are separated from A83543 as a mixture (H:J ratio = 58:42) which can be separated by analytical high performance liquid chromatography (HPLC), as described infra . The structures assigned to A83543H and J are based on ¹H and ¹³C NMR studies of the A83543 H:J mixture in acetone-d₆. The NMR spectra resemble and were compared with those of A83543A. The major changes in the H and J spectra center around the rhamnose sugar. Components H and J have only two OCH₃'s in that sugar. In component H, H-1′ is shifted about .1 δ in the ¹H NMR spectrum and 3 δ in the ¹³C NMR spectrum (4.81 and 99.68, respectively). These shifts are due to the absence of methyl on the methoxy at the 2′ position. The shifts in component J correspond to a similar absence of the methyl on the methoxy at the 3′ position. Characteristics of A83543A Pseudoaglycone Molecular Weight: 590 Empirical Formula: C₃₃H₅₀O₉ UV (EtOH) λmax: 243 nm (ε 10,300) IR (CHCl₃): ν (lactone) 1724; (conjugated ketone 1652; multiple peaks for C-H vibrations around 3017; multiple peaks for C-O vibrations around 1140 cm ⁻¹ (see fig. 3) FD-MS: see Fig. 7 EI-MS: see Fig. 11 Characteristics of A83543D Pseudoaglycone Molecular Weight: 604 Empirical Formula: C₃₄H₅₂O₉ The A83543 components can be separated from each other using one of the following analytical HPLC systems: SYSTEM I Column: ODS, 3 µ, 4.5 x 50 mm (IBM) Solvent: CH₃OH:CH₃CN:H₂O (2:2:1) Flow Rate: 1.0 mL/min Detection: UV at 245 nm Temperature: Room Temperature Component Retention Time (min) A 8.50 B 6.15 C 3.92 D 11.47 SYSTEM II Column: 4.6 x 100 mm, ODS (AQ-301, S-5; YMC, Inc., Mt. Freedom, NJ) Solvents: CH₃OH:CH₃CN:0.05% NH₄OAc(H₂O) (A) 35:35:30 - pH 7.8 (B) 45:45:10 - pH 6.7 Flow Rate: 2.0 mL/min Run time: 35 min Detection: UV, 250 nm Gradient: 10% B to 25% B in 20 min; to 50% B in 30 min Component Retention Time (min) A 22.62 A-pseudoaglycone 7.27 B 12.65 C 10.62 D 25.47 E 19.22 F 16.30 G 18.92 H 16.30 J 17.50 SYSTEM III Column: 4.6 x 100 mm, ODS (AQ-301, S-5; YMC, Inc., Mt. Freedom, NJ) Solvents: CH₃OH:CH₃CN:0.05% NH₄OAc(H₂O) 35:35:30 - pH 6.0 Flow Rate: 2.0 mL/min Run time: 10 min Detection: UV, 250 nm Component Retention Time (min) F 4.32 H 3.42 J 3.42 The A83543 components are not soluble in water, but are soluble in solvents such as methanol, ethanol, dimethylformamide, dimethyl sulfoxide, acetonitrile, acetone and the like. A83543 and individual components, such as A83543A, A83543B, A83543C, A83543D, A83543E, A83543F, A83543G, A83543H, and A83543J can react to form various salts. All such forms of these compounds are part of this invention. A83543 salts are useful, for example, for separating and purifying A83543. In addition, some salts have an improved solubility in water. A83543 salts are prepared using standard procedures for salt preparation. For example, A83543 can be neutralized with an appropriate acid to form an acid addition salt. The acid addition salts are particularly useful. Representative suitable salts include those salts formed by standard reactions with both organic and inorganic acids such as, for example, sulfuric, hydrochloric, phosphoric, acetic, succinic, citric, lactic, maleic, fumaric, cholic, pamoic, mucic, glutamic, camphoric, glutaric, glycolic, phthalic, tartaric, formic, lauric, stearic, salicylic, methanesulfonic, benzenesulfonic, sorbic, picric, benzoic, cinnamic and like acids. In discussions of utility, the term "A83543 compound" preferably denotes a member selected from the group consisting of A83543, individual components A83543A, A83543B, A83543C, A83543D, A83543E, A83543F, A83543G, A83543H, and A83543J and their acid addition salts. Fermentation product A83543 is produced by culturing a strain of the novel microorganism Saccharopolyspora spinosa selected from NRRL 18395, NRRL 18537, NRRL 18538 and NRRL 18539, or an A83543-producing mutant thereof, under submerged aerobic conditions in a suitable culture medium until a recoverable amount of A83543 is produced. As those skilled in fermentation processes will recognize, the ratio of the components in A83543 will vary, depending upon the fermentation conditions used to produce it. In general, A83543 contains about 85-90% A83543A, about 10-15% A83543D and minor amounts of A83543B, C, E, F, G, H and J and A83543A pseudoaglycone. Individual components such as A83543A, A83543B, A83543C, A83543D, A83543E, A83543F, A83543G, A83543H, and A83543J and A83543A pseudoaglycone can be separated and isolated as described infra . Thus, another aspect of this invention is a process for producing an A83543 compound which comprises cultivating a Saccharopolyspora spinosa strain selected from NRRL 18395, NRRL 18357, NRRL 18538 or NRRL 18539, or an A83543-producing mutant thereof, in a culture medium containing assimilable sources of carbon, nitrogen, and inorganic salts under submerged aerobic fermentation conditions until a recoverable amount of A83543 is produced. Individual components can then be isolated by techniques known per se in the art. This invention also relates to a biologically purified culture of the microorganism Saccharopolyspora spinosa selected from NRRL 18395, NRRL 18537, NRRL 18538 or NRRL 18539, or an A83543-producing mutant thereof. These microorganisms are useful because they produce A83543. For convenience in the discussions which follow, the strains have been given the following designations: A83543.1, A83543.3, A83543.4 and A83543.5. Culture A83543.1 was obtained by chemical mutation of a culture (A83543) isolated from a soil sample collected from the Virgin Islands. Cultures A83543.3, A83543.4 and A83543.5 were obtained from derivatives of the A83543.1 culture by chemical mutations. Cultures A83543.1, A83543.3, A83543.4 and A83543.5 have been deposited and made part of the stock culture collection of the Midwest Area Northern Regional Research Center, Agricultural Research Service, United States Department of Agriculture, 1815 North University Street, Peoria, Illinois, 61604, from which they are available to the public under the following accession numbers: NRRL No. Strain No. 18395 A83543.1 18537 A83543.3 18538 A83543.4 18539 A83543.5 Taxonomic studies of culture A83543.1 were carried out by Frederick P. Mertz of the Lilly Research Laboratories. Based on these studies, the microorganisms A83543.1, A83543.3, A83543.4 and A83543.5 are classified as members of a new species of the genus Saccharopolyspora , which is called Saccharopolyspora spinosa sp . nov . This classification is based on direct laboratory comparisons and examination of published descriptions of similar species. Methods Used The methods followed were those recommended by the International Streptomyces Project (ISP) for the characterization of Streptomyces species [E. B. Shirling and D. Gottlieb, "Methods for Characterization of Streptomyces Species," Int. J. Syst. Bacteriol. 16 :313-340 (1966)] and those recommended for the characterization of Nocardia species by R. E. Gordon, D. A. Barnett, J. E. Handerhan and C. H. Pang, " Nocardia coeliaca , Nocardia autotrophica , and the Nocardin Strain ", Int. J. Syst. Bacteriol. 24 (1), 54-63 (1974). ISCC-NBS Centroid Color Charts, standard sample No. 2106 (National Bureau of Standards, 1958, U.S. Department of Commerce, Washington, D.C.) were used to assign color names to the reverse side and to aerial hyphae. Morphology was studied using an optical light microscope and a scanning electron microscope (SEM). The isomer of diaminopimelic acid (DAP) and the carbohydrates in hydrolysates of whole cells were established by the chromatographic methods of Becker et al. [B. Becker, M. P. Lechevalier, R. E. Gordon and H. E. Lechevalier, "Rapid Differentiation between Nocardia and Streptomyces by Paper Chromatography of Whole-cell Hydrolysates," Appl. Microbiol. 12 , 421-423 (1964)] and of Lechevalier and Lechevalier [M. P. Lechevalier and H. Lechevalier, "Chemical Composition as a Criterion in the Classification of Aerobic Actinomycetes," Int. J. Syst. Bacteriol. 20 , 435-443 (1970)]. Phospholipids were determined by the procedure of M. P. Lechevalier and H. Lechevalier [in A University Laboratory Approach , Dietz and Thayer (eds.), Society for Industrial Microbiology Special Publication No. 6, Arlington, VA, pp. 227-233 (1980)]. Menaquinone composition was determined by following the procedures of R. M. Kroppenstedt [in Chemical Methods in Bacterial Systematics , M. Goodfellow and D. E. Minnikin (eds.), 1985, pp. 173-196] and M. D. Collins ( ibid. , pp. 267-285). Resistance to antibiotics was measured by padding antibiotic sensitivity discs onto the surface of seeded ISP No. 2 agar plates. Starch hydrolysis was determined by testing for the presence of starch with iodine on ISP No. 4 (inorganic salts-starch) agar plates. Fatty acid analysis was done using the HP 5898A Microbial Identification System [see L. Miller and T. Berger, "Bacterial Identification by Gas Chromatography of Whole Cell Fatty Acids," Hewlett-Packard Application Note 228-41, 8pp. (1985)]. Fatty acid methyl esters were made from lyophilized whole cells grown under identical conditions. Principal component analysis was two dimensional and computer generated. Units of measurement in the principal component plot (shown in Figure 12) are standard deviations. Mycolic acids were determined by the methods proposed by Minnikin [D. E. Minnikin, I. G. Hutchinson and A. B. Caldicott, "Thin-Layer Chromatography of Methanolysates of Mycolic Acid-Containing Bacteria," J. Chromatography 188 , 221-233 (1980)]. Cultural Characteristics Culture A83543.1 grew well on both complex and defined media. The culture produced aerial mycelia on all the media used. The aerial spore-mass color was predominantly light yellowish-pink, but was white on a number of the media. The reverse side was yellow to yellow-brown. No distinctive pigmentation was present. A soluble brown pigment was released into the medium in some media. The cultural characteristics are summarized in Table VI. Morphological Characteristics Culture A83543.1 produced an extensive substrate mycelium that fragmented in liquid fermentation. No fragmentation was observed when the culture was grown on agar media. White circular colonies, 8-10 mm in diameter with raised center and yellow-brown reverse color, were observed when the culture was plated on ISP medium 1. Well-formed aerial hyphae were present on most of the media. The aerial hyphae were segmented into long chains of spores arranged as hooks and open loops. Spirals were also observed, but they were short and incomplete. The general morphology was Rectus-flexibilis (RA). Aerial hyphae had a distinctive bead-like appearance, with many empty spaces in the spore chain. This feature demonstrated that a spore sheath encased the spore chain. This spore sheath was covered with very distinctive spines. The spines were approximately 1 µm long and were rounded on the end. The spore shape was oblong and averaged approximately 1.1 x 1.5 µm in size. The spore-chain length was well over 50 spores. No zig-zag characteristics, sclerotia, sporangia or motile cells were observed. Physiological Characteristics Culture A83543.1 produced acid from the following carbohydrates: adonitol, D-arabinose, erythritol, fructose, glucose, glycerol, mannitol, mannose, ribose and trehalose. The culture did not produce acid from: L-arabinose, cellobiose, cellulose, dextrin, dulcitol, ethanol, galactose, glycogen, inositol, inulin, lactose, maltose, melizitose, melebiose, α-methyl-D-glucoside, raffinose, L-rhamnose, salicin, sorbitol, L-sorbose, sucrose, xylitol or xylose. Growth was observed with galactose, maltose and melizitose, but no acid was produced from these carbohydrates. Culture A83543.1 used the following organic acids as sodium salts: acetate, butyrate, citrate, formate, lactate, malate, propionate, pyruvate and succinate. The culture did not use benzoate, mucate, oxalate or tartrate. A83543.1 decomposed allantoin, calcium malate, casein, elastin, hippurate, hypoxanthine, testosterone, L-tyrosine and urea. It was unable to decompose adenine, esculin, guanine, starch or xanthine. A83543.1 produced catalase, phosphatase, urease and H₂S. It liquefied gelatin and reduced nitrate. It was not resistant to lysozyme and did not produce melanoid pigments. It neither peptonized nor hydrolyzed skim milk. A83543.1 tolerated levels of NaCl up to and including 11%. It was unable to survive 50°C for 8 hours, but grew at temperatures between 15° and 37°C. A83543.1 was resistant to cephalothin (30 µg), penicillin G (10 units) and rifampin (5 µg). It was sensitive to bacitracin (10 units), gentamicin (10 µg), lincomycin (2 µg), neomycin (30 µg), oleandomycin (15 µg), streptomycin (10 µg), tetracycline (30 µg), tobramycin (10 µg) and vancomycin (30 µg). Cell-Wall Analysis Hydrolyzed whole cells of A83543.1 contained meso -diaminopimelic acid. Diagnostic sugars in the whole-cell extracts were galactose and arabinose. Thus, A83543.1 has a Type IV cell-wall pattern and a Type A sugar pattern (Lechevalier and Lechevalier, supra ). The cells do not contain mycolic acids. Phospholipid determinations on the whole cells indicated the presence of phosphatidyl choline and cardiolipin. No phosphatidyl ethanolamine was detected. Thus, A83543.1 has a Type PIII phospholipid pattern [M. P. Lechevalier, A. E. Stern and H. A. Lechevalier, "Phospholipids in the Taxonomy of Actinomycetes ," in Actinomycetes, Zbl. Bakt. Suppl. 11 , K. P. Schaal and G. Pulverer (eds), Gustav Fischer Verlag, New York, 1981]. The major menaquinone detected was MK-9(H₄). A minor amount of MK-9(H₆) was observed. Phage Plating A number of Streptomycete , Saccharopolyspora and Amycolatopsis phages were plated on A83543.1. No plaques were observed. Identity of A83543.1 As discussed, supra , culture A83543.1 has a type IV cell-wall pattern and a type A whole-cell sugar pattern. The following thirteen genera have this pattern of cell chemistry: Nocardia , Rhodococcus , Corynebacterium , Caseobacter , Mycobacterium , Faenia ( Micropolyspora ), Pseudonocardia , Saccharomonospora , Saccharopolyspora , Actinopolyspora , Amycolata , Amycolatopsis and Kibdelosporangium . These genera are distinguished by the presence or absence of mycolic acids, by fatty acid composition, and by phospholipid and menaquinone types. Faenia , Pseudonocardia and Saccharopolyspora have chemotaxonomic characteristics identical to those of A83543.1, but these genera differ from A83543.1 in morphological and cultural properties. The genus Faenia ( Micropolyspora ) has smooth spores and short spore chains which are borne on both aerial and substrate hyphae. Its aerial hyphae are sparse and white in color. It is a thermophile which grows at 60°C. A83543.1 has none of these properties and thus differs from Faenia . The genus Pseudonocardia has spores on both aerial and substrate hyphae. It is distinguished by acropetal budding and blastospores. It has a characteristic zig-zag morphology of the hyphae. The hyphae have been described as articulated but non-septate [see A. Henssen and D. Schafer, "Amended Description of the Genus Pseudonocardia Henssen and Description of a New Species Pseudonocardia spinosa Schafer," Int. J. Syst. Bacteriol. 21 :29-34 (1971)]. It grows very slowly. Fragmentation is absent or rarely observed. A83543.1 has none of these properties. The genus Saccharopolyspora is characterized by a spore sheath and a distinctive bead-like appearance of the spore chain. This feature is very prominent in A83543.1. Fragmentation has also been observed with the genus Saccharopolyspora . The type species, S . hirsuta , was isolated from sugar-cane bagasse. The parent culture from which A83543.1 was obtained was isolated from a sugar mill. Since A83543.1 has many properties of this genus, it is, therefore, considered to be a strain of Saccharopolyspora . The only validly published species in the genus Saccharopolyspora are S . erythraea and S . hirsuta . Known subspecies are S . hirsuta subsp. taberi and S . hirsuta subsp. kobensis . A83543.1 differs from these strains in either aerial and reverse color or in production of soluble pigments. Biochemical similarity was measured by constructing a table of similarity coefficients based on as many biochemical measurements as possible. The coefficient of Jaccard Sj and the simple matching coefficient S sm were used [see W. Kurylowicz, A. Paszkiewicz, W. Woznicka, W. Kurzatkowski and T. Szulga, "Numerical Taxonomy of Streptomycetes," Polish Medical Publishers, Warsaw, 1975, p. 37]. Table VII summarizes these similarity coefficients. Similarity Coefficients For A83543.1 and Saccharopolyspora species Culture S sm Sj A83543.1 100 100 S . hirsuta subsp. taberi 68 57 S . hirsuta subsp. kobensis 67 60 S . erythraea 63 54 S . hirsuta 55 50 Fatty acid analysis of A83543.1 and the known Saccharopolyspora species showed that each has both saturated and branched-chain fatty acids. The fatty acid composition of A83543.1 is similar, but not identical, to that of the other species. Table VIII compares the fatty acid compositions of A83543.1 and the known Saccharopolyspora species. Principal component analysis of the fatty acid compositions shown in Table VIII show sufficient scattering to suggest the cultures are all distinct species within the same genus. The principal component plot of the Table VIII data is presented in Figure 12. Table IX compares the physiological characteristics of A83543.1 to those of the existing Saccharopolyspora species and subspecies. The comparisons made indicate that A83543.1 differs sufficiently from the previously described species of Saccharopolyspora to be a new species of Saccharopolyspora for which the name Saccharopolyspora spinosa has been selected. The name spinosa reflects the spiny spore ornamentation of this species. The A83543.3, A83543.4 and A83543.5 strains are sufficiently similar macroscopically to the A83543.1 strain to be classified as strains of Saccharopolyspora spinosa . The four strains differ in the amount of A83543 they produce. The A83543.3 strain produces approximately four-fold more A83543 than the A83543.1 strain; and the A83543.4 and A83543.5 strains produce approximately eight to nine-fold more than the A83543.1 strain produces. As is the case with other organisms, the characteristics of the A83543-producing cultures of this invention, Saccharopolyspora spinosa NRRL 18395, NRRL 18537, NRRL 18538 and NRRL 18539, continue to be subject to variation. Thus, mutants of these strains may be obtained by physical and chemical methods known in the art. For example, other strains can be obtained by treatment with chemicals such as N-methyl-N′-nitro-N-nitrosoguanidine. Natural and induced mutants of the Saccharopolyspora spinosa NRRL 18395, NRRL 18537, NRRL 18538 and NRRL 18539 strains which retain the characteristic of producing a recoverable amount of A83543 are part of this invention. The culture medium used to grow the Saccharopolyspora spinosa cultures can be any one of a number of media. For economy in production, optimal yield, and ease of product isolation, however, certain culture media are preferred. Thus, for example, preferred carbon sources in large-scale fermentation are glucose and maltose, although ribose, xylose, fructose, galactose, mannose, mannitol, soluble starch, potato dextrin, methyl oleate, oils such as soybean oil and the like can also be used. Preferred nitrogen sources are cottonseed flour, peptionized milk and digested soybean meal, although fish meal, corn steep liquor, yeast extract, enzyme-hydrolyzed casein, beef extract, and the like can also be used. Among the nutrient inorganic salts which can be incorporated in the culture media are the customary soluble salts capable of yielding zinc, sodium, magnesium, calcium, ammonium, chloride, carbonate, sulfate, nitrate and like ions. Essential trace elements necessary for the growth and development of the organism should also be included in the culture medium. Such trace elements commonly occur as impurities in other substituents of the medium in amounts sufficient to meet the growth requirements of the organism. Usually, if foaming is a problem, small amounts (i.e., 0.2 ml/L) of an antifoam agent such as polypropylene glycol) may be added to large-scale fermentation media. In the case of the A83543-producing cultures, however, conventional defoamers inhibit A83543 production. Foaming can be controlled by including soybean oil or pluronic L-101 (BASF) in the medium (1-3%). Additional oil may be added if foaming develops. The percentage of a particular A83543 component may be varied by media changes. For example, adding valine or isobutyric or propionic acids increases the percentage of A83543D produced. For production of substantial quantities of A83543, submerged aerobic fermentation in stirred bioreactors is preferred. Small quantities of A83543 may be obtained by shake-flask culture. Because of the time lag in production commonly associated with inoculation of large bioreactors with the spore form of the organism, it is preferable to use a vegetative inoculum. The vegetative inoculum is prepared by inoculating a small volume of culture medium with the spore form or mycelial fragments of the organism to obtain a fresh, actively growing culture of the organism. The vegetative inoculum is then transferred to a larger bioreactor. The vegetative inoculum medium can be the same as that used for larger fermentations, but other media are also suitable. A83543 is produced by the A83543-producing organisms when grown at temperatures between about 24° and about 33°C. Optimum temperatures for A83543 production appear to be about 28-30°C. As is customary in submerged aerobic culture processes, sterile air is blown into the vessel from the bottom while the medium is stirred with conventional turbine impellors. In general, the aeration rate and agitation rate should be sufficient to maintain the level of dissolved oxygen at or above 35%, and preferably at or above 50%, of air saturation with an internal vessel pressure of 0.34 atmospheres. Production of the A83543 components can be followed during the fermentation by testing extracts of the broth. HPLC, using a system as described in Example 1, is a useful assay for this purpose. Following their production under submerged aerobic fermentation conditions, the A83543 components can be recovered from the fermentation medium by methods used in the art. The A83543 produced during fermentation of the A83543-producing organism occurs in both the mycelia and the broth. A83543 appears to be lipophilic. Thus, if a substantial amount of oil is used in the fermentation, whole broth extraction is more efficient. If only small amounts of oil are used, the major portion of the A83543 is in the mycelia. In that case, more efficient recovery of A83543 is accomplished by initially filtering the medium to separate the broth from the mycelial mass (the biomass). A83543 can be recovered from the biomass by a variety of techniques. A preferred technique involves washing the separated biomass with water to remove remaining broth, mixing the biomass with a polar solvent in which A83543 is soluble, e.g. methanol or acetone, separating and concentrating the solvent, extracting the concentrate with a non-polar solvent and/or adsorbing it onto a reverse-phase silica gel adsorbent such as RP-C8 or RP-C18 or a high porous polymer like HP-20 and the like. The active material is eluted from the adsorbent with a suitable solvent such as, for example, acetonitrile:methanol mixtures containing small amounts of THF. A83543 can be separated into individual components A83543A, A83543B, A83543C, A83543D, A83543E, A83543F, A83543G, A83543H, and A83543J and A83543A pseudoaglycone by similar procedures. A preferred separation procedure involves reverse-phase silica-gel (C₁₈ or C₈) chromatography. Alternatively, the culture solids, including medium constitutents and mycelium, can be used without extraction or separation, but preferably after removal of water, as a source of A83543. For example, after production of A83543, the whole fermentation broth can be dried by lyophilization, by drum-drying, or by azeotropic distillation and drying. The dried broth can then be used directly, e.g. by mixing it directly into feed premix. Insecticide and Miticide Activity The compounds of this invention are useful for the control of insects and mites. The present invention is also directed, therefore, to methods for inhibiting an insect or mite which comprises applying to the locus of the insect or mite an insect- or mite-inhibiting amount of an A83543 compound. The A83543 compounds show activity against a number of insects and mites. More specifically, the compounds show activity against Southern armyworm, which is a member of the insect order Lepidoptera. Other typical members of this order are codling moth, cutworms, clothes moths, Indianmeal moth, leaf rollers, corn earworm, cotton bollworm, European corn borer, imported cabbageworm, cabbage looper, pink bollworm, bagworms, Eastern tent caterpillar, sod webworm and fall armyworm. The compounds also show activity against cotton aphid, which is a member of the insect order Homoptera. Other members of the Homoptera include leafhoppers, planthoppers, pear psylla, apple sucker, scale insects, whiteflies and spittle bugs, as well as a number of other host-specific aphid species. In addition, the A83543 compounds show activity against stable flies, blow flies and mosquitoes, which are members of the insect order Diptera. Another typical member of this order is the common house fly. The A83543 compounds are useful for reducing populations of insects and mites, and are used in a method of inhibiting an insect or mite population which comprises applying to a locus of the insect or mite an effective insect- or mite-inactivating amount of an A83543 compound. The "locus" of insects or mites refers to the environment in which the insects or mites live or where their eggs are present, including the air surrounding them, the food they eat, or objects which they contact. For example, plant-ingesting insects or mites can be controlled by applying the active compound to plant parts which the insects or mites eat or inhabit, particularly the foliage. It is contemplated that the compounds might also be useful to protect textiles, paper, stored grain, or seeds by applying an active compound to such substances. The term "inhibiting an insect or mite" refers to a decrease in the number of living insects or mites or to a decrease in the number of viable insect or mite eggs. The extent of reduction accomplished by a compound depends, of course, upon the application rate of the compound, the particular compound used, and the target insect or mite species. At least an insect-inactivating or mite-inactivating amount should be used. The terms "insect-inactivating amount" and "mite-inactivating amount" are used to describe the amount which is sufficient to cause a measurable reduction in the treated insect or mite population. Generally an amount in the range from about 1 to about 1000 ppm (or 0.01 to 1 kg/ha) of active compound is used. In one preferred embodiment, the present invention is directed to a method for inhibiting a susceptible insect of the order Lepidoptera which comprises applying to a plant an effective insect-inactivating amount of an A83543 compound in accordance with the present invention. Another preferred embodiment of the invention is directed toward a method for inhibiting biting flies of the order Diptera in animals which comprises administering an effective pest-inhibiting amount of an A83543 compound orally or parenterally to the animal. MITE/INSECT SCREEN The A83543 compounds were tested for miticidal and insecticidal activity in the following mite/insect screen. Each test compound was formulated by dissolving the compound in an acetone/alcohol (1:1) mixture containing 23 g of "Toximul R" (sulfonate/nonionic emulsifier blend) and 13 g of "Toximul S" (sulfonate/nonionic emulsifier blend) per liter. These mixtures were then diluted with water to give the indicated concentrations. Two-spotted spider mites ( Tetranychus urticae Koch) and cotton or melon aphids ( Aphis gossypii Glover) were introduced on squash cotyledons and allowed to establish on both leaf surfaces. Other plants in the same treatment pot were left uninfested. The leaves were then sprayed with 5 mL of test solution using a DeVilbiss atomizing sprayer at 10 psi. Both surfaces of the leaves were covered until runoff and then allowed to dry for one hour. Two uninfested leaves were then excised and placed into a Petri dish containing Southern armyworm ( Spodoptera eridania Cramer). Additional insects were evaluated using similar formulations and evaluation procedures, with the exceptions noted. After standard exposure periods, percent mortality was evaluated. Results are reported in the tables infra . The following abbreviations are used: Term Insect/Mite Scientific Name BW boll weevil Anthonomus grandis CA cotton aphid Aphis gossypii CBW cotton bollworm Heliothis zea CLH corn leafhopper Dalbulus maidis CRW Southern corn rootworm Diabrotica undecimpunctata howardi SAW Southern armyworm Spodoptera eridania SM twospotted spider mite Tetranychus urticae Percent Control of Neonate CBW Larvae by A83543 Components Component PPM 0.5 1 5 10 A 55 (d) 100 (a) 100 (a) 100 (a) B 35 (e) 80 (c) 100 (a) 100 (a) C 5 (g) 85 (bc) 93 (bc) 100 (a) D 58 (d) 90 (ac) 100 (a) 100 (a) E 28 (ef) 58 (d) 100 (a) 100 (a) F 0 (g) 0 (g) 0 (g) 95 (ab) G 0 (g) 0 (g) 20 (f) 80 (c) Activity of A83543A Against Adult BW Treatment Rate (ppm) % Inhibition Topical 1.00 0.00 5.00 20.00 10.00 20.00 50.00 100.00 100.0 100.00 Table XV compares the effectiveness and persistence of A83543A treatment with that of methomyl in outdoor pot tests against Southern armyworm. Efficacy of A83543 Against Spodoptera eridania in Outdoor Pot Tests Treatment Rate (ppm) Days After Treatment 1 5 7 14 A83543A 63 100 88 90 5 A83543A 125 100 88 95 90 A83543A 250 100 95 100 100 A83543A 500 100 100 100 100 methomyl 63 100 20 15 0 methomyl 125 100 35 40 20 methomyl 250 100 85 45 40 methomyl 500 100 90 85 60 None 0 0 0 0 0 None 0 0 0 0 0 Table XVI summarizes the LC 50's, the lethal concentrations at which test compound inhibits 50% of test insect or mite, exhibited by A83543A as compared with that exhibited by the known insecticide methomyl. LC 50'S for A83543 Target LC 50 (ppm) A83543 methomyl CRW 15 6 Grain Aphid >250 5.4 SM 150 71.4 SAW/Foliar 1.3 11.4 CBW/Contact 0.6 14.5 The A83543 components were active against yellow fever mosquito (Aedes aegypti ) larvae in standard in vitro mosquito larvicide tests. Tables XVII and XVIII summarize the activity of the components in these tests. In Vitro Activity of A83543B and A83543C Against First Instar Mosquito Larvae A83543 Component Minimal Inhibitory Concentration (mcg/mL) A 0.016, 0.03l B 0.016 C 0.031 Activity of A83543 Components vs. Fourth Instar Mosquito Larvae a Component Percent Control A 60,70 B 60 C 60 D 30 E 80 F 0 G 0 Field Trials A83543A was evaluated in field trials. In these trials, A83543A showed activity against imported cabbage worm and cabbage looper on sprouting broccoli and against a mixture of soybean loopers (75%) and fall armyworm (25%) on soybean. Insecticidal Compositions The compounds of this invention are applied in the form of compositions, which are also part of this invention. These compositions comprise an insect-or mite-inactivating amount of an A83543 compound and a phytologically acceptable inert carrier. The active component, i.e. the A83543 compound, may be present as 1) a single A83543 component, 2) a mixture of two or more components, 3) the separated A83543 mixture or 4) A83543 together with the dried portion of the fermentation medium in which it is produced, i.e. the as crude, dried fermentation broth. The compositions are either concentrated formulations which are dispersed in water for application, or are dust or granular formulations which are applied without further treatment. The compositions are prepared according to procedures and formulae which are conventional in the agricultural chemical art, but which are novel and important because of the presence of one or more of the compounds of this invention. The dispersions in which the compounds or crude dried material are applied are most often aqueous suspensions or emulsions prepared from concentrated formulations of the compounds or crude material. Such water-soluble, water-suspendable or emulsifiable formulations are either solids (usually known as wettable powders) or liquids (usually known as emulsifiable concentrates or aqueous suspensions). Wettable powders, which may be compacted to form water dispersible granules, comprise an intimate mixture of the active compound, an inert carrier and surfactants. The concentration of the active compound is usually from about 1%, preferably 10%, to about 90% by weight. The inert carrier is usually chosen from among the attapulgite clays, the montmorillonite clays, the diatomaceous earths, or the purified silicates. Effective surfactants, comprising from about 0.5% to about 10% of the wettable powder, are found among the sulfonated lignins, the condensed naphthalenesulfonates, the naphthalenesulfonates, the alkylbenzenesulfonates, the alkyl sulfates, and non-ionic surfactants such as ethylene oxide adducts of alkyl phenols. Emulsifiable concentrates of the compounds comprise a convenient concentration of a compound, such as from about 50 to about 500 grams per liter of liquid, equivalent to about 10% to about 50%, dissolved in an inert carrier which is either a water miscible solvent or a mixture of water-immiscible organic solvent and emulsifiers. Useful organic solvents include aromatics, especially the xylenes, and the petroleum fractions, especially the high-boiling naphthalenic and olefinic portions of petroleum such as heavy aromatic naphtha. Other organic solvents may also be used, such as the terpenic solvents including rosin derivatives, aliphatic ketones such as cyclohexanone, and complex alcohols such as 2-ethoxyethanol. Suitable emulsifiers for emulsifiable concentrates are chosen from conventional nonionic surfactants, such as those mentioned above. Aqueous suspensions comprise suspensions of water-insoluble compounds of this invention, dispersed in an aqueous vehicle at a concentration in the range from about 5% to about 50% by weight. Suspensions are prepared by finely grinding the compound, and vigorously mixing it into a vehicle comprised of water and surfactants chosen from the same types discussed above. Inert ingredients, such as inorganic salts and synthetic or natural gums, may also be added to increase the density and viscosity of the aqueous vehicle. It is often most effective to grind and mix the compound at the sametime by preparing the aqueous mixture and homogenizing it in an implement such as a sand mill, ball mill, or piston-type homogenizer. The compounds may also be applied as granular compositions, which are particularly useful for applications to the soil. Granular compositions usually contain from about 0.5% to about 10% by weight of the compound, dispersed in an inert carrier which consists entirely or in large part of clay or a similar inexpensive substance. Such compositions are usually prepared by dissolving the compound in a suitable solvent and applying it to a granular carrier which has been pre-formed to the appropriate particle size, in the range of from about 0.5 to 3 mm. Such compositions may also be formulated by making a dough or paste of the carrier and compound, and crushing and drying to obtain the desired granular particle size. Dusts containing the compounds are prepared simply by intimately mixing the compound in powdered form with a suitable dusty agricultural carrier, such as kaolin clay, ground volcanic rock and the like. Dusts can suitably contain from about 1% to about 10% of the compound. It is equally practical, when desirable for any reason, to apply the compound in the form of a solution in an appropriate organic solvent, usually a bland petroleum oil, such as the spray oils, which are widely used in agricultural chemistry. Insecticides and miticides are generally applied in the form of a dispersion of the active ingredient in a liquid carrier. It is conventional to refer to application rates in terms of the concentration of active ingredient in the carrier. The most widely used carrier is water. The compounds of the invention can also be applied in the form of an aerosol composition. In such compositions the active compound is dissolved or dispersed in an inert carrier, which is a pressure-generating propellant mixture. The aerosol composition is packaged in a container from which the mixture is dispensed through an atomizing valve. Propellant mixtures comprise either low-boiling halocarbons, which may be mixed with organic solvents, or aqueous suspensions pressurized with inert gases or gaseous hydrocarbons. The actual amount of compound to be applied to loci of insects and mites is not critical and can readily be determined by those skilled in the art in view of the examples provided. In general, concentrations of from 10 ppm to 5000 ppm of compound are expected to provide good control. With many of the compounds, concentrations of from 100 to 1000 ppm will suffice. For field crops, such as soybeans and cotton, a suitable application rate for the compounds is about 0.01 to 1 kg/ha, typically applied in 5-50 gal/A of spray formulation. The locus to which a compound is applied can be any locus inhabited by an insect or mite, for example, vegetable crops, fruit and nut trees, grape vines, and ornamental plants. Because of the unique ability of mite eggs to resist toxicant action, repeated applications may be desirable to control newly emerged larvae, as is true of other known acaricides. Ectoparasiticide Activity The A83543 compounds are also active against members of the insect order Diptera. Tables XIX - XXI summarize in vitro studies with A83543A and A83543D. In vitro Efficacy of A83543A Against Black Blow Fly Larvae Dose Level (ppm) Activity 100 100 50 100 25 100 10 100 5 100 2 100 1 95 0.5 60 0.25 25 In vitro Efficacy of A83543A Against Adult Stable Fly Dose Level (ppm) 24 Hour Activity¹ 48 Hour Activity 100 100 100 50 100 100 25 100 100 10 100 90 5 80 100 2 70 90 1 10 60 0.5 0 10 In in vivo tests, the A83543 compounds showed systemic insecticidal activity in guinea pigs and sheep against larval blow fly and adult stable fly with no obvious signs of toxicity. Representative compounds A83543A and A83543D have been tested in laboratory and target animals to determine the scope of activity. The following tests are illustrative. Guinea Pig Systemic Test Adult guinea pigs are used in this test system. The compounds to be tested are dissolved in aqueous polyvinyl pyrrolidone or in polyethylene glycol 200, and an appropriate amount of the solution is administered either orally or by intraperitoneal injection. Various doses of the compound are used, as set forth in the tables infra . Blood is drawn from the guinea pigs at 30 minutes except as noted, and the samples of blood are centrifuged. Dental wicks are saturated with the blood serum, and then exposed in Petri dishes to adult stable flies; blow fly larvae are exposed in test tubes. After 24 and 48 hours, the insects are examined, and the number of dead are counted. The results of the tests are recorded as the percent of the insects which are killed. Sheep Systemic Test Tests are carried out in sheep using the test method described supra in the Guinea Pig Test description. Test compound is administered by intraperitoneal or intravenous injection or intraruminally; blood samples are drawn at 24 hours in these tests. The results of guinea pig and sheep systemic tests using A83543A and A83543D are summarized in Tables XXII - XXV. A83543A did not show anthelmintic activity in sheep treated by a single intraperitoneal or intraruminal dose of 50 mg/kg of body weight, or in mice experimentally infected with the intestinal nematode Nematospiroides dubius , when administered orally by a single gavage at 500 mg/kg. In in vitro tests against the sheep intestinal nematode Haemonchus contortus , A83543A killed 30% of the worms at a concentration of 100 ppm. Ectoparasiticidal Methods This invention also relates, therefore, to a method of controlling a population of insect ectoparasites which consume blood of a host animal which comprises administering to the host animal an effective amount of an A83543 compound. Insect ectoparasites include insect and acarina parasites. Administration to the animal may be by the dermal, oral or parenteral routes. Parasitic insects and acarina include species that are bloodsucking as well as flesh eating and are parasitic during all of their life cycle or only part of their life cycle, such as only the larval or only the adult stage. Representative species include the following: horse fly Tabanus spp. stable fly Stomoxys calcitrans black fly Simulium spp. horse sucking louse Haematopinus asini mange mite Sarcoptes scabiei scab mite Psoroptes equi horn fly Haematobia irritans cattle biting louse Bovicola bovis shortnosed cattle louse Haematopinus eurysternus longnosed cattle louse Linognathus vituli tsetse fly Glossina spp. cattle follicle mite Demodex bovis cattle tick Boophilus microplus and B . decoloratus Gulf Coast tick Amblyomma maculatum Lone Star tick Amblyomma americanum ear tick Otobius megnini Rocky Mountain wood tick Dermacentor andersoni screwworm fly Cochliomyia hominivorax assassin bug Reduvius spp. mosquito Culiseta inornata brown ear tick Rhipicephalus appendiculatus African red tick Rhipicephalus evertsi bont tick Amblyomma sp. bont legged tick Hyalomma sp. hog louse Haematopinus suis chigoe Tunga penetrans body louse Haematopinus ovillus foot louse Linognathus pedalis sheep ked Melophagus ovinus sheep scab mite Psoroptes ovis greenbottle fly Phaenicia sericata black blow fly Phormia regina secondary screw-worm Cochliomyia macellaria sheep blow fly Phaenicia cuprina bed bug Cimex lectularius Southern chicken flea Echidnophaga gallinacea fowl tick Argas persicus chicken mite Dermanyssus gallinae scalyleg mite Knemidokoptes mutans depluming mite Knemidokoptes gallinae dog follicle mite Demodex canis dog flea Ctenocephalis canis American dog tick Dermacentor variabilis brown dog tick Rhipicephalus sanguineus The method of the invention may be used to protect economic and companion animals from ectoparasites. For example, the compounds may beneficially be administered to horses, cattle, sheep, pigs, goats, dogs, cats and the like, as well as to exotic animals such as camels, llamas, deer and other species which are commonly referred to as wild animals. The compounds may also beneficially be administered to poultry and other birds, such as turkeys, chickens, ducks and the like. Preferably, the method is applied to economic animals, and most preferably to cattle and sheep. The rate, timing, and manner of effective application will vary widely with the identity of the parasite, the degree of parasiticidal attack, and other factors. Applications can be made periodically over the entire lifespan of the host, or for only a peak season of parasitic attack. In general, ectoparasite control is obtained with topical application of liquid formulations containing from about 0.00005 to 95.0% of compound, preferably up to 5%, and most preferably up to about 1% of compound. Effective parasite control is achieved at administration rates of from about 5 to about 100 mg/kg. The compounds are applied to host animals by conventional veterinary practices. Usually, the compounds are formulated into ectoparasiticidal compositions which comprise a compound and a physiologically-acceptable carrier. For example, liquid compositions may be simply sprayed on the animals for which ectoparasiticidal control is desired. The animals may also treat themselves by such devices as back rubbers, which may contain the toxicant compound in a cloth, for example, which the animal may walk against and contact. Dip tanks are also employed to administer the active agent to the host animal. The present compounds display systemic ectoparasiticidal activity. The compounds have the ability to permeate the tissues of a host animal to which one of the compounds has been administered. Insect parasites which then consume blood or other living tissues of the host animal are thereby killed. The compounds are administered by dermal, oral or percutaneous routes. This invention also relates to ectoparasiticidal compositions comprising a physiologically-acceptable inert carrier and an A83543 compound. These compositions may be prepared by methods known in the art, for example by dissolving the compound in one of many physiologically-acceptable adjuvants or diluents. Oral administration may be performed by mixing the compound in the animals' feed or drinking water, or by administering dosage forms such as tablets, capsules, boluses, or implants. Percutaneous administration is conveniently accomplished by subcutaneous, intraperitoneal and intravenous injection of an injectable formulation. The compounds can be formulated for oral administration in the usual forms, such as drenches, tablets, or capsules. Such compositions, of course, require orally-acceptable inert carriers. The compounds can also be formulated as an injectable solution or suspension, for subcutaneous, dermal, intraruminal, intraperitoneal, intramuscular, or intravenous injection. In some applications, the compounds are conveniently formulated as one component of a standard animal feed. In this embodiment, it is usual to formulate the present compound first as a premix in which the compound is dispersed in a liquid or particulate solid carrier. The premix can contain from about 2 to 250 grams of compound per pound. The premix is in turn formulated into the ultimate feed by conventional mixing. Since ectoparasitic attack generally takes place during a substantial portion of the host animal's lifespan, it is preferred to administer the compounds of the present invention in a form to provide sustained release over a period of time. Conventional procedures include the use of a matrix which physically inhibits dissolution, where the matrix is a waxy semisolid such as the vegetable waxes or a high molecular weight polyethylene glycol. A good way to administer the compounds is by means of a sustained-action bolus, such as those of Laby, U.S. Patent 4,251,506, and Simpson, British Patent 2,059,767. For such a bolus, the compound would be encapsulated in a polymeric matrix such as that of Nevin, U.S. Patent 4,273,920. Sustained release of the compounds of the present invention can also be achieved by the use of an implant such as from a silicone-containing rubber. In order to illustrate more fully the operation of this invention, the following examples are provided: EXAMPLE 1 A83543 HPLC Assay Method The following analytical HPLC method is useful for monitoring the fermentation for production of A83543: Centrifuge a sample of the whole broth, decant and remove the supernatant. Add enough methanol to the biomass to return the sample to the original volume, mix, and allow the mixture to stand a minimum of fifteen minutes. Centrifuge and filter the supernatant through a 0.45 µ filter. Alternatively, the whole broth can be extracted with acetonitrile (1:4 broth:solvent) or acetone. HPLC System : Column Support: 8-x 100-mm column, silica gel-4µ spherical C₁₈ (Nova C18, Waters) Mobile Phase: CH₃CN/MeOH/H₂O (45/45/10) containing 0.05% ammonium acetate Flow Rate: 4 mL/min Detection: UV at 250 nm Retention Times: A83543A - 3.6 - 3.7 min A83543D - 4.4 - 4.5 min EXAMPLE 2 Preparation of A83543 with Culture A83543.1 A. Shake-flask Fermentation The culture Saccharopolyspora spinosa NRRL 18395, either as a lyophilized pellet or as a suspension maintained in liquid nitrogen, is used to inoculate a vegetative medium having composition A or B (medium B is preferred for large scale production): VEGETATIVE MEDIUM A Ingredient Amount (%) Trypticase soy broth 3.0 Yeast extract 0.3 MgSO₄.7H₂O 0.2 Glucose 0.5 Maltose 0.4 Deionized water q.s. 1 liter No pH adjustment VEGETATIVE MEDIUM B Ingredient Amount (%) Enzyme-hydrolyzed casein 3.0 Yeast extract 0.3 MgSO₄.7H₂O 0.2 Glucose 1.0 Deionized water q.s. 1 L pH 6.2, adjust to 6.5 with NaOH Slants or plates can be prepared by adding 2.5% agar to vegetative seed medium A or B. The inoculated slant is incubated at 30°C. for from about 10 to 14 days. The mature slant culture is scraped with a sterile tool to loosen the spores and remove and macerate the mycelial mat. About one-fourth of the loosened spores and culture growth thus obtained is used to inoculate 50 mL of a first-stage vegetative seed medium. Alternatively, the first-stage medium may be inoculated from a liquid nitrogen ampoule. When the culture is maintained in liquid nitrogen, ampoules are prepared using equal volumes of vegetative culture (48-72 hr incubation, 30°C) and suspending medium. The suspending medium contains lactose (100 g), glycerol (200 mL) and deionized water (q.s. to 1 L). A liquid nitrogen ampoule is used to inoculate 100 mL of vegetative medium in 500-mL Erlenmeyer flasks (or 50 mL medium in 250-mL flasks). The cultures are incubated at 30°C for 48 hours on a shaker orbiting in a two-inch (5.08 cm) circle at 250 rpm. The incubated culture (5% v/v inoculum) is used to inoculate 100 mL of a production medium having the following composition: PRODUCTION MEDIUM I Ingredient Amount (%) Glucose 4 Vegetable protein, partially hydrolyzed enzymatically 1.5-3 Cottonseed flour 1.0 CaCO₃ (reagent or technical grade) 0.3 Soybean oil 1.0 Tap water q.s. 1 liter (Presterilization pH adjusted to 7.0 with NaOH) The inoculated production medium is incubated in 500-mL Erlenmeyer flasks at 28-30°C. for 6 to 8 days on a shaker orbiting in a two-inch circle at 250 rpm. B. Stirred Bioreactor Fermentation In order to provide a larger volume of inoculum, 10 mL of incubated first-stage medium, prepared as described in Section A, is used to inoculate 400 mL of a second-stage vegetative medium having the same composition as that of the first-stage vegetative medium. This second-stage medium is incubated in a 2-L wide-mouth Erlenmeyer flask for about 48 hours at 30°C. on a shaker orbiting in a two-inch circle at 250 rpm. Incubated second-stage vegetative medium (2 L) thus prepared is used to inoculate 80 to 115 liters of sterile production medium, prepared as described in Section A. Additional soybean oil is added to control foaming, if needed. The inoculated production medium is allowed to ferment in a 165-L stirred bioreactor for 5 to 8 days at a temperature of 28°C. The airflow and agitator speed in the stirred vessel are computer controlled to maintain a dissolved oxygen level at or above 50% of air saturation. EXAMPLE 3 Isolation of A83543A, B, C and D Fermentation broth (225 liters), prepared as described in Example 2, was filtered using a filter aid (1% Hyflo), and the separated biomass was washed with water (∼ 50 L). The biomass was then agitated with methanol (∼ 100 L) for about one hour and filtered. The methanol filtrate was concentrated to a volume of about 1 liter. The concentrate was extracted three times with diethyl ether (1 L each). The combined ether extracts were concentrated to a volume of about 200 mL. A portion of the concentrate (8 mL) was chromatographed on a silica gel column (RP-8 Lobar, size B, E.M. Science, a Division of E.M. Industries, Inc.). This procedure was repeated for a total of 12 runs (cycles). The instrumental set-up and execution procedure to perform the preparative chromatography in an "Autoprep" mode is described as follows: A complete "Autoprep" HPLC system is comprised of three Rainin Rabbit HPX pumps, one pressure module, one Gilson Model 20 1B HPLC fraction collector, one ISCO-V⁴ absorbance detector and one Apple Macintosh Plus computer. The complete system is arranged according to instructions given in the Dynamax HPLC Method Manager manual from Rainin Instrument Company, Inc. The "Autoprep" HPLC configuration takes advantage of system automation to permit preparative separations to be run repetitively under virtually identical conditions with virtually identical results. Collecting and pooling corresponding fractions from multiple runs provides chromatographic capacity without the need for a large column. Two solvent mixtures (A) and (B) are used in the isocratic mode at a flow rate of 8.0 mL/min. Solvent Systems Solvent Amount (mL) A B CH₃OH 95 100 CH₃CN 95 100 H₂O 10 - The isocratic mixture used contains 60% of solvent B. The runtime for each cycle is 28.0 minutes. The eluates from the first 16 minutes of each run are discarded. The following eluates are collected in 6 time-factions, 2 minutes (16 mL) each. The automatically combined fractions from each of the 12 cycles resulted in 6 final fractions (chromatographic cuts). The presence of the active A83543 compounds is determined by analyzing each final fraction for mosquito larvae activity and also by analytical HPLC. The active fractions are then combined according to their activity and HPLC profiles and are further purified, using the same "Autoprep" HPLC and solvent system, but with a high resolution, 21.4-mm x 25-cm preparative column (Rainin Dynamax), prepacked with 8 µ C-18 reversed phase silica gel, to give A83543 components A, B, C and D. Factors A and D crystallize from CH₃OH/H₂O. The FD-MS of A83543C is shown in figure 5; and the FAB-MS of A83543B is shown in figure 9 of the drawings. EXAMPLE 4 Purification of A83543A and D Fermentation broth (10 L) was prepared as described in Example 2 Sect. A, except that 1) 200 mL of production medium was used in 1-L flasks; 2) soybean oil was omitted from the production medium; and 3) incubation was at 30° for 4-6 days. The broth was filtered. The filtrate, containing 4 mcg of A83543A/ mL and no detectable quantities of A83543B, C, or D/mL, was discarded. The biomass was washed with water and extracted for one hour with methanol. The extract (7 L) contained 72 mcg of A83543A/mL and 7 mcg of A83543D/mL. The methanol extract was concentrated to a volume of 5 L, and added to HP-20 resin (150 mL, Mitsubishi Chemical Industries, Ltd., Japan) in water (2 L). This mixture was stirred for one hour. The HP-20 resin mixture was then placed in a glass column. The initial effluent and the eluate using methanol:water (1:1, 1 L) were not active. The second eluate using methanol:water (7:3, 1 L) contained trace quantities of A83543A. The following eluate using methanol (1 L) contained the A83543A and A83543D activity. The methanol eluate was concentrated and combined with 2 similar fractions from other work-ups and concentrated to dryness. The residue was dissolved in 75 mL of methanol:THF (4:1) and precipitated by addition into 10 volumes of acetonitrile. The mixture was filtered, and the filtrate was concentrated to dryness. The residue was dissolved in methanol (25 mL) and applied to a 5.5-x 90-cm column of LH-20 Sephadex (Pharmacia LKB Biotechnology Inc., U.S.A.), prepared in methanol, collecting and analyzing 125 25-mL fractions, using the HPLC procedure described in Example 1. Fractions containing the desired compounds were combined and concentrated. The residue was dissolved in methanol (10 mL) and applied to a 41.1-mm x 25-cm preparative column prepacked with 8µ C-18 reversed phase silica gel (Rainin Dynamax). The column was conditioned in methanol: acetonitrile:water (37.5:37.5:25). After sample application, the column was developed using a 180-min linear gradient of the following solvents: Solvent System Solvent Amount (mL) A B CH₃OH 37.5 45 CH₃CN 37.5 45 H₂O 25 10 The gradient was run from 100% A to 100% B, collecting 25-mL fractions. Fractions containing A83543A were pooled, concentrated to dryness, dissolved in t -BuOH (5 mL) and lyophilized to give 778 mg of pure A83543A. The IR spectrum of A83543A in CHCl₃ is shown in figure 1. The FD-MS spectrum of A83543A is shown in figure 4; the FAB-MS spectrum of A83543A is shown in figure 8 [FAB dispersent-dithiothreitol: dithioerythritol (5:1); and the EI-MS spectrum of A83543A is shown in figure 10. Fractions containing A83543D were combined with D-containing fractions from 6 similar separations and were concentrated and chromatographed as described supra , using the same column but different solvents. The column was conditioned in methanol:acetonitrile: water(40:40:20). The solvent systems used to develop the column in a 180-min linear gradient operation were: Solvent Systems Solvent Amount (mL) A B CH₃OH 40 95 CH₃CN 40 95 H₂O 20 10 Fractions containing A83543D were combined and concentrated. The residue was dissolved in t -BuOH (5 mL) and lyophilized to give 212 mg of A83543D. The IR spectrum of A83543D in CHCl₃ is shown in figure 2; and the FD-MS spectrum of A83543D is shown in figure 6. EXAMPLE 5 Isolation of A83543 Components E,F,G,H and J and the Pseudoaglycone of A Fermentation broth (8 L), prepared using procedures similar to those described in Example 2, was treated as described in Example 4. Fractions from the LH-20 Sephadex column containing the desired compounds were combined with corresponding fractions from similar fermentations. Since components E, F, G, H, J and the pseudoaglycone of A are produced in very small quantities, numerous fermentations are required to provide sufficient quantities for further purification. A pool of minor factors, prepared in this manner and containing approximately 1.6 grams of solid material, was applied to an HPLC column (Rainin Dynamax) prepacked with 8 micron C-18 reversed phase silica gel (ODS), as described in Example 4. The column was conditioned in CH₃OH:CH₃CN:H₂0 (75:75:50), and the gradient was run from 100% of solvent (A) to 50% (B): with the following solvent systems: Solvent Systems Solvent Amount (%) A B CH₃OH 75 95 CH₃CN 75 95 H₂O 50 10 collecting 25-mL fractions. The following fractions were pooled: Pool Fractions 1 31 - 44 2 45 - 63 3 64 - 69 4 70 - 80 5 81 - 130 6 131 - 160 A portion of pool 5 (100 mL) was concentrated to a residue, dissolved in methanol (1 mL) and applied to a 21.4-mm x 250-mm HPLC column (Rainin Dynamax), as described in Example 3. The column was conditioned using solvent system (A) of the following solvent systems: Solvent Systems Solvent Amount (%) A B CH₃OH 30 95 CH₃CN 30 95 H₂O(1N NH₄OAC,pH5.0) 40 -- H₂O -- 10 and developed using a 120-minute linear gradient from 100% solvent (A) to 50% of solvent (B), collecting 15-mL fractions at 7.5 mL/min. Elution was continued at 50% (B) for an additional 60 minutes. The following fractions were pooled: Pool Fraction Component 1 37 F 2 38 - 48 E 3 52 - 63 B,G 4 65 - 70 H,J These pools were combined with pools from other chromatographic runs using similar starting materials. The combined pools were further purified using column chromatography, as described supra ; desalted on HP-20 resins, using standard techniques; and concentrated and lyophilized to give the following components: Component Quantity (mg) Mol. Wt IR E 249 717 Fig. 13 F 4 717 Fig. 14 G 104 731 Fig. 15 H,J 87 717 Fig. 16 Pseudo A 288 590 Fig. 3 The FD-MS and EI-MS spectra of A83543A pseudoaglycone are shown in Figures 7 and 11, respectively. EXAMPLE 6 Preparation of A83543 with Culture A83543.3 A. Shake-flask Fermentation Using procedures like those of Example 2, Section A, the culture Saccharopolyspora spinosa NRRL 18537 is cultivated in shake flasks, but using vegetative medium C as follows: VEGETATIVE MEDIUM C Ingredient Amount Enzyme-hydrolyzed casein 30.0 g Yeast extract 3.0 g MgSO₄.7H₂O 0.2 g Glucose 10.0 g Glycerol 20 mL Deionized water q.s. 1 L pH 6.6, adjust to 7.0 with NaOH B. Stirred Bioreactor Fermentation Liquid nitrogen ampoules of the culture are prepared as described in Example 2, using the general procedures of Sect. B. One ampoule is used to inoculate a first-stage vegetative culture (50 mL of medium C in 250-mL flasks), which is incubated for about 48-72 hours. Incubated first-stage culture is used to inoculate (10-mL inoculum) a second-stage culture (400 mL of medium C in 2-L flasks), which is incubated for about 48 hrs. The incubated second-stage culture (5 L) is used to inoculate a production medium (115 L) having the following composition: PRODUCTION MEDIUM II Ingredient Amount (g/L) Glucose 80 Peptonized milk 20 Cottonseed flour 20 CaCO₃ 5 Methyl oleate 30 mL/L Tap water q.s. to 1 L The inoculated production medium is allowed to ferment in a 165-L stirred bioreactor for about 8 to 10 days, or longer, at a temperature of 30°C. Dissolved oxygen (DO) levels are regulated by computerized systems set to maintain the DO level above 50% of air saturation as described in Example 2, Section B. EXAMPLE 7 Preparation of A83543 with Culture A83543.5 A. Shake-flask Fermentation Using procedures like those of Example 2, Section A, the culture Saccharopolyspora spinosa NRRL 18539 is cultivated in shake flasks, using vegetative medium B. B. Stirred Bioreactor Fermentation Liquid nitrogen ampoules of the culture are prepared as described in Example 2, using the general procedures of Sect. B. One ampoule is used to inoculate a first-stage vegetative culture (50 mL of medium B in 250-mL flasks), which is incubated for 48 to 72 hours. Incubated first-stage culture is used to inoculate (10-mL inoculum) a second-stage culture (400 mL of medium B in 2-L flasks), which is incubated for about 48 hr. The incubated second-stage culture (2 L) is used to inoculate a production medium (115 L) having one of the following compositions: PRODUCTION MEDIUM III Ingredient Amount (g/L) Glucose 80 Vegetable protein, partially hydrolyzed 20 enzymatically Cottonseed flour 10 CaCO₃ 5 Methyl oleate 30 mL/L Tap water q.s. to 1 L PRODUCTION MEDIUM IV Ingredient Amount (Percent) Glucose 8 Cottonseed flour 3 Peptionized Milk 2 Corn steep liquor 1 CaCO₃ (tech. grade) 0.5 Methyl oleate 3.0 Tap water q.s. to 1 L The inoculated production medium is allowed to ferment in a 165-L stirred bioreactor for about 8 to 10 days, or longer, at a temperature of 30°C. DO levels are regulated as described in Example 6. EXAMPLE 8 Preparation of A83543 with Culture A83543.4 Using procedures like those of Examples 2 and 7, the culture Saccharopolyspora spinosa NRRL 18538 is cultivated but using vegetative medium B and production medium III. EXAMPLE 9 Fermentation broth is prepared as described in Example 8. A83543 is separated from the broth as follows: 1. Add an equal volume of acetone to the broth and filter, using a ceramic filter or filter aid with a filter press. 2. Adjust the filtrate broth to pH 10. 3. Add ethyl acetate (¼-½ the volume of the broth). 4. Recover the ethyl acetate extract by decanting off the immiscible aqueous portion; and concentrate the ethyl acetate extract to ½ volume by vacuum. 5. Extract the concentrated ethyl acetate solution with aqueous 0.1 M tartaric acid (½ volume); and separate the phases. 6. Remove the soluble ethyl acetate from the aqueous phase by vacuum (about 5%) evaporation. Concentrate the aqueous solution, using a reverse osmosis operation. 7. Adjust the concentrated aqueous solution to pH 10-11 with sodium hydroxide. 8. Separate the precipitate by filtration; wash with water; and dry under vacuum to give A83543. EXAMPLE 10 A83543A Pseudoaglycone A sample of A83543 containing mostly component A (ca 100 mg) was dissolved into methanol (50 mL), water (2 mL) and concentrated HCl (3 mL). This solution was concentrated to dryness at 50°C. The residue was treated twice with diethyl ether (200 mL each), and the insoluble material was discarded. The combined ether solutions containing the crude pseudoaglycone were concentrated to dryness. The residue was dissolved in methanol (20 mL) and purified, using the AUTOPREP-HPLC system described in Example 3, to give 20 mg of pure A83543A pseudoaglycone. EXAMPLE 11 A83543D pseudoaglycone is prepared from A83543D, using a procedure similar to that described in Example 10. EXAMPLE 12 The following formulations are typical of the insecticidal compositions useful in this invention. A. Aqueous Suspension A-83543A 12.5% "Tergitol TMN-6" (nonionic surfactant) 1.0% "Zeosyl 200" (silica) 1.0% "AF-100" (silicon based antifoam agent) 0.2% Xanthan solution (2%) 10.0% "Makon 10" (10 moles ethyleneoxide nonylphenol surfactant) 9.0% Tap water 66.3% B. Emulsifiable Concentrate A83543D 12.4% "Exxon 200" (naphthalene solvent) 83.6% "Toximul H" (nonionic/anionic surfactant blend) 2.0% "Toximul D" (nonionic/anionic surfactant blend) 2.0% EXAMPLE 13 The following exemplary compositions illustrate the sort of formulations used to practice the method of the present invention. A. Feed Premix A83543A 10% Rice hulls 85 Light mineral oil 5 B. Feed Premix A83543E 25% Alfalfa meal 60 Powdered clay 5 Molasses 10 C. Suspension A83543A 30% Naphthalenesulfonate salt 5 Nonionic surfactant 5 Fumed silica 1 Water 59 D. Drip-On Solution A83543A 20% Nonionic surfactant 0.8 Propylene glycol 15 Water 64.2 E. Drip-On Suspension A83543B 10 Nonionic surfactant 1 Light mineral oil 89 F. Injectable Solution A83543A 15% Propylene glycol 85 G. Injectable Suspension A83543C 25% Propylene glycol 15 Water 60 H. Injectable Suspension A83543D 30% Polyvinylpyrrolidone 2 Water 68

Claims: Claims for the following Contracting States : AT, BE, CH, DE, FR, GB, GR, IT, LI, LU, NL, SE An A83543 compound of formula 1 : wherein R is H or a group selected from R² is R¹, R³, R⁵ and R⁶ are hydrogen or methyl; R₄ is methyl or ethyl; or an acid addition salt thereof where R is other than hydrogen. An A83543 compound of formula 1 as claimed in claim 1 wherein R, R¹, R³, R⁴ R⁵ and R⁶ are present in one of the following combinations: R R₁ R³ R⁴ R⁵ R⁶ (a) Me H Et Me Me (b) Me H Et Me Me (c) Me H Et Me Me (a) Me Me Et Me Me (a) Me H Me Me Me (a) H H Et Me Me (d) Me H Et Me Me (a) Me H Et H Me (a) Me H Et Me H H Me H Et Me Me H Me Me Et Me Me H Me H Me Me Me H H H Et Me Me H Me H Et H Me H Me H Et Me H or an acid addition salt thereof when R is other than hydrogen. Compound A83543A of Claim 2 wherein: R is an (a) group; R¹, R⁵ and R⁶ = CH₃; R³ = H; and R⁴ = ethyl; or an acid addition salt thereof. Compound A83543D of claim 2 wherein: R is an (a) group; R¹, R³, R⁵ and R⁶ = CH₃; and R⁴ = ethyl; or an acid addition salt thereof. Fermentation product A83543, comprising A83543A and A83543D, which is produced by culturing a Saccharopolyspora spinosa strain selected from NRRL 18395, NRRL 18537, NRRL 18538 or NRRL 18539, or an A83543-producing mutant thereof, under submerged aerobic conditions in a suitable culture medium until a recoverable amount of A83543 is produced. A process for producing an A83543 compound as claimed in any one of claims 1 to 4 which comprises cultivating a Saccharopolyspora spinosa strain selected from NRRL 18395, NRRL 18537, NRRL 18538 or NRRL 18539, or an A83543-producing mutant thereof, in a culture medium containing assimilable sources of carbon, nitrogen, and inorganic salts under submerged aerobic fermentation conditions until a recoverable amount of A83543 is produced. A biologically purified culture of Saccharopolyspora spinosa selected from NRRL 18395, NRRL 18537, NRRL 18538, or NRRL 18539, or an A83543-producing mutant thereof. An insecticide or miticide composition comprising an A83543 compound as claimed in any one of claims 1 to 4 and a phytologically-acceptable carrier. A method for inhibiting an insect or mite which comprises applying to the locus of the insect or mite an A83543 compound as claimed in any one of claims 1 to 4. An ectoparasiticidal composition comprising a physiologically-acceptable inert carrier and an A83543 compound as claimed in any one of claims 1 to 4. Claims for the following Contracting State: ES An insecticide or miticide composition comprising as an active ingredient from 1 to 90% by weight of an A83543 compound of formula 1 : wherein R is H or a group selected from R² is R¹, R³, R⁵ and R⁶ are hydrogen or methyl; R₄ is methyl or ethyl; or a phytologically acceptable salt thereof where R is other than hydrogen; associated with one or more phytologically acceptable carriers or diluents therefor. An insecticide- or miticide formulation as claimed in claim 1 wherein the A83543 compound of formula is one in which R, R¹, R³, R⁴ R⁵ and R⁶ are present in one of the following combinations: R R₁ R³ R⁴ R⁵ R⁶ (a) Me H Et Me Me (b) Me H Et Me Me (c) Me H Et Me Me (a) Me Me Et Me Me (a) Me H Me Me Me (a) H H Et Me Me (d) Me H Et Me Me (a) Me H Et H Me (a) Me H Et Me H H Me H Et Me Me H Me Me Et Me Me H Me H Me Me Me H H H Et Me Me H Me H Et H Me H Me H Et Me H or an acid addition salt thereof when R is other than hydrogen. An insecticide- or miticide formulation as claimed in any one of claims 1 or 2 wherein the A83543 compound is one wherein: R is an (a) group; R¹, R⁵ and R⁶ = CH₃; R³ = H; and R⁴ = ethyl, or an acid addition salt thereof. An insecticide- or miticide formulation as claimed in any one of claims 1 or 2 wherein the A83543 compound is one wherein: R is an (a) group; R¹, R³, R⁵ and R⁶ = CH₃; and R⁴ = ethyl; or an acid addition salt thereof. A method for inhibiting an insect or mite which comprises applying to the locus of the insect or mite an effective amount of an A83543 compound, or a phytologically accepable salt thereof, as defined in any one of claims 1 to 4. An ectoparasiticidal formulation comprising as an active ingredient from 0.00005 to 95% by weight of an A83543 compound of formula 1 as defined in claim 1 or a physiologically acceptable salt thereof associated with one or more physiologically-acceptable inert carriers or diluents therefor. A process for producing an A83543 compound of formula 1 as defined in any one of claims 1 to 4 which comprises cultivating a Saccharopolyspora spinosa strain selected from NRRL 18395, NRRL 18537, NRRL 18538 or NRRL 18539, or an A83543-producing mutant thereof, in a culture medium containing assimilable sources of carbon, nitrogen, and inorganic salts under submerged aerobic fermentation conditions until a recoverable amount of A83543 is produced. A biologically purified culture of Saccharopolyspora spinosa selected from NRRL 18395, NRRL 18537, NRRL 18538, or NRRL 18539, or an A83543-producing mutant thereof.

Inventors: BOECK LAVERNE DWAINE, CHIO HANG, EATON TOM EDWARD, GODFREY OTIS WEBSTER JR, MICHEL KARL HEINZ, NAKATSUKASA WALTER MITSUO, YAO RAYMOND CHE-FONG, BOECK, LAVERNE DWAINE, CHIO, HANG, EATON, TOM EDWARD, GODFREY, OTIS WEBSTER, JR., MICHEL, KARL HEINZ, NAKATSUKASA, WALTER MITSUO, YAO, RAYMOND CHE-FONG
Applicants: DOWELANCO
Application year: 1989
Main classification: C07H17
IPC codes: A01N43, C12P19, C12P1, C07H17, C12R1, C12N1