US 2013 O156740A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2013/015674.0 A1 Leland (43) Pub. Date: Jun. 20, 2013 (54) BIO-PESTCIDE METHODS AND (52) U.S. Cl. COMPOSITIONS CPC ................ A0IN 63/04 (2013.01); A0IN 63/02 (71) Applicant: Novozymes Biologicals Holdings A/S, (2013.01) Bagsvaerd (DK) USPC ......................................................... 424/93.5 (72) Inventor: Jarrod E. Leland, Blacksburg, VA (US) (57) ABSTRACT (73) Assignee: NOVOZYMES BIOLOGICALS - 0 HOLDINGS A/S, Bagsvaerd (DK) The present invention is directed to the combination of bio pesticide and at least one exogenous cuticle degrading (21) Appl. No.: 13/719,624 enzymes (e.g., a protease, chitinase, lipase and/or cutinase) (22) Filed: Dec. 19, 2012 for controlling (preventing or eliminating) pests. The use of an exogenous cuticle degrading enzyme increases the effi Related U.S. Application Data cacy of the biopesticide by increasing the speed and/or effi (60) Provisional application No. 61/577,224, filed on Dec. ciency of infestation of the pest resulting in faster or more 19, 2011. effective killing or disabling of the pest by the biopesticide. The present invention accordingly provides methods for con Publication Classification trolling a pest comprising treating a pest habitat with a com (51) Int. Cl. bination of pesticidally effective amounts of at least one bio AOIN 63/04 (2006.01) pesticide and at least one exogenous cuticle degrading AOIN 63/02 (2006.01) enzyme. Pest control compositions are also described. Patent Application Publication Jun. 20, 2013 US 2013/015674.0 A1 Effect of Protease and Metarhizium on Mortality at Varying Concentrations O 1. 2 3 A. 5 6 7 8 s Days s0% Savinase No Met x&x 0.1% Savinase No Met NSw1% Savinase No Met xxx 10% Savinase No Met &O% Savinase 3E6 Met O.1% Savinase 3E6 Met w x 1%, Savinase 3E6 Met ·.10% Savinase 3E6 Met FIG. 1 US 2013/0156740 A1 Jun. 20, 2013 BO-PESTCIDE METHODS AND effective amounts of at least one entomopathogenic fungus COMPOSITIONS and at least one exogenously applied cuticle degrading enzyme. The present invention is also directed to an insecti CROSS-REFERENCE TO RELATED cide composition comprising insecticidally effective APPLICATIONS amounts of at least one entomopathogenic fungus and at least one exogenous cuticle degrading enzyme. 0001. This application claims the benefit under 35 U.S.C. 0007. In another aspect, the present invention is directed to 119 of U.S. provisional application No. 61/577,224 filed Dec. a method of controlling an Acari infestation comprising treat 19, 2011, the contents of which are fully incorporated herein ing a pest habitat with a combination of pesticidally effective by reference. amounts of at least one acaripathogenic fungus and at least BACKGROUND one exogenously applied cuticle degrading enzyme. The present invention is also directed to a pesticide composition 0002 Pests, such as insects, Acari (mites and ticks) and comprising pesticidally effective amounts of at least one nematodes, are a major problem for the agriculture industry, acaripathogenic fungus and at least one exogenous cuticle limiting productivity, often significantly. Although chemical degrading enzyme. pesticides are used to control pests, excessive use of chemical 0008. The present invention is also directed to a method of pesticides leaves residues in Soil, water and air and also has controlling a nematode infestation comprising treating a adverse effects on the non-target organisms and the ecologi nematode habitat with a combination of nematicidally effec cal balance. In addition, pests can develop resistance to tive amounts of at least one nematopathogenic fungus and at chemical pesticides, limiting their effectiveness and applica least one exogenously applied cuticle degrading enzyme. The tion. Public concern overpotential health hazards of chemical present invention is also directed to a nematicide composition pesticides and the increase in cost of chemical pesticides has comprising nematicidally effective amounts of at least one also led to the exploration of more eco-friendly pest manage nematopathogenic fungus and at least one exogenous cuticle ment tactics. degrading enzyme. 0003 Biopesticides have been developed for use as an 0009. An entomopathogenic fungus may also have nem alternative, or in some cases as a Supplement, to chemical atopathogenic properties, and vice versa. Alternatively, at pesticides. Biopesticides are living organisms (e.g., fungi and least one entomopathogenic fungus and at least one nemato bacteria) that intervene in the life cycle of pests (by killing or pathogenic fungus may be used in combination as ingredients disabling the pest). Examples of biopesticides include the of pest treatment composition. In an embodiment, the present entomopathogenic fungus Metarhizium anisopliae, which invention provides a method of controlling a pest comprising has been registered as a bio-insecticide for the control of treating a pest habitat with a combination of pesticidally insect pests in the United States and many other countries. effective amounts of at least one entomopathogenic fungus, at Metarhizium anisopliae has been reported to infect many least one nematopathogenic fungus and at least one exog insect types including Subterranean termites (Reticulitermes enously applied cuticle degrading enzyme, wherein the at and Coptotermes spp.), corn rootworms (Diabrotica spp). least one entomopathogenic fungus and the at least one nem black vine weevils (Otiorhynchus sulcatus), citrus root wee atopathogenic fungus may be the same fungus or a different vils (Diaprepes abbreviatus), Japanese beetles (Popifia fungus. japonica), and European chafers (Rhizotrogus maialis). 0004 As natural agents, biopesticides offer more eco BRIEF DESCRIPTION OF THE DRAWING friendly solutions for controlling pests and/or for for use in combination with chemical pesticide. However, one major 0010 FIG. 1. is a graph that illustrates the effect of pro drawback of the use of biopesticides is in their efficacy com tease and Metarhizium on mortality at varying concentra pared to chemical pesticides. There is a need in the art for tions. biopesticides having greater efficacy for the replacement or Supplementation of chemical pesticides. DETAILED DESCRIPTION OF THE INVENTION 0011. A fungal pesticide is employed in the present inven BRIEF SUMMARY OF THE INVENTION tion to protect a habitat from pests (such as, an insect, an 0005. The present invention is directed to the combination Acari, and/or a nematode infestation) so as to prevent, elimi of at least one biopesticide and at least one exogenously nate or reduce a pest infestation in a habitat. As used herein, applied cuticle degrading enzymes (e.g., a protease, chitinase, the term “fungal pesticide” means a fungal organism that is lipase and/or cutinase) for use in controlling pests. The use of pathogenic to a target pest, Such as, an insect, Acari or a an exogenously applied cuticle degrading enzyme increases nematode. the efficacy of the biopesticide. The present invention accord 0012. As used herein, a “habitat” may be any area or ingly provides methods and compositions for controlling a environment where a pest lives or is able to live, that is, any pest comprising treating a pest habitat with a combination of area or environment that is infested or susceptible to infesta pesticidally effective amounts of at least one biopesticide and tion by a pest. at least one exogenously applied cuticle degrading enzyme. 0013 The habitat may be a plant, soil, or water, as well as, Pests which may be treated according to the present invention commercial or residential structures, storage containers (e.g., include, for example, insects, Acari (Such as, mites and ticks) shipping containers), and commercial products (e.g., food and/or nematodes (and, accordingly, the biopesticide may be products) and product packaging. The habitat may be agri used as an insecticide, Acaricide, and/or nematicide). cultural fields, orchards, greenhouses, gardens, lawns, orna 0006. In one aspect, the present invention is directed to a mental plants, or trees. method of controlling an insect infestation comprising treat 0014. The at least one fungal pesticide and the least one ing an insect habitat with a combination of insecticidally cuticle degrading enzyme are applied to the habitat in the US 2013/0156740 A1 Jun. 20, 2013 vicinity of the target pest, such as, e.g., in agriculture, on the The term "isolated” means the enzyme is in a form or envi Surface of the plants to be protected (e.g., as a foliar applica ronment which does not occur in nature, that is, the enzyme is tion), as a seed coating, and/or to the soil, using conventional at least partially removed from one or more or all of the techniques. As used herein, “vicinity’ means a location effec naturally occurring constituents with which it is associated in tive to result in treatment of the pest by immediate or eventual nature. Thus, although enzymes produced endogenously by contact with the pest. The application or treating process will the fungal pesticide will impact efficacy, an isolated enzyme vary depending on the habitat. does not encompass an enzyme endogenously produced by 0015 The term “entomopathogenic’ means that the fun the fungal pesticide during treatment of apest in the processes gal pesticide is pathogenic to at least one targetinsect. As used of the present invention. An isolated enzyme may be present herein, "entomopathogenic fungus' is a fungus that can act as