WO2018037361A1

WO2018037361A1 - Miticidal, insecticidal and nematocidal compositions containing plants extracts from bulnesia sp and humulus sp

Info

Publication number: WO2018037361A1
Application number: PCT/IB2017/055094
Authority: WO
Inventors: James JIMÉNEZ MARTINEZ; Juan Fernando Gil Romero; Camilo CHACÓN; Gladis JARAMILLO
Original assignee: Ecoflora Agro Sas
Priority date: 2016-08-23
Filing date: 2017-08-23
Publication date: 2018-03-01

Abstract

The present disclosure relates to biopesticide compositions comprising a Bulnesia sp. plant extract (e.g., an extract from Bulnesia sarmientoi) and a Humulus sp. plant extract (e.g., an extract from Humulus lupulus). In some aspects, the Bulnesia sp. plant extract and the Humulus sp. plant extract act synergistically as miticides. Also provided are methods of manufacture, formulations, and kits comprising the disclosed compositions. The disclosed compositions can be use control, treat, or prevent pest infestations (e.g., by mites, insects, nematodes, etc.) in plants and in animals, including humans, in need thereof.

Description

MITICIDAL, INSECTICIDAL AND NEMATOCIDAL COMPOSITIONS CONTAINING

PLANTS EXTRACTS FROM BULNESIA SP AND HUMULUS SP

BACKGROUND

[0001] Mites are among the most diverse and successful of all the invertebrate groups.

Many species live as parasites on plants and animals. It is estimated that 48,200 species of mites have been described. Many mites which have been well studied are parasitic on plants and animals. Dust mites feed mostly on dead skin and hair shed from humans. Insects may also be infested by parasitic mites. For example, Varroa destructor, attaches to the body of the honey bee, and Acarapis woodi (family Tarsonemidae) lives in the tracheae of honey bees.

[0002] Some of the plant pests include the so-called spider mites (family Tetranychidae), thread-footed mites (family Tarsonemidae), and the gall mites (family Eriophyidae). Among the species that attack animals are members of the sarcoptic mange mites (family Sarcoptidae), which burrow under the skin. Demodex mites (family Demodicidae) are parasites that live in or near the hair follicles of mammals, including humans.

[0003] Some species of mites can colonize humans directly, act as vectors for disease transmission, or cause or contribute to allergenic diseases. Mites which colonize human skin are the cause of several types of skin itchy rashes, such as grain itch, grocer's itch, and scabies.

[0004] Dust mites cause several forms of allergic diseases, including hay fever, asthma and eczema, and are known to aggravate atopic dermatitis. House dust mites are usually found in warm and humid locations, including beds. It is thought that inhalation of mites during sleep exposes the human body to some antigens that eventually induce

hypersensitivity reaction.

[0005] Many chemicals which possess insecticidal properties have been found to possess little or no effect in killing mites, whereas other chemicals are toxic to mites in the adult stage but not in the egg stage and hence must be applied at very short intervals in order to control the mites from cycle to cycle. Accordingly, there is a need to provide effective miticide compositions that will control mites from the egg to the adult stage.

[0006] Frequent treatment of plants with many chemicals which are effective as miticides results in most instances in serious damage to the plant foliage. Most of the chemicals which have been found effective for the control of mites in plants, shrubs and trees, tend to burn the plant. In many instances substantial injury to the foliage is not tolerable. For example, in the control of mites on ornamental plants, only minor degrees of leaf burn can be tolerated. In other instances, for example in the case of citrus fruits, the injury to the fruit caused by the treating chemical frequently gives rise to a down-grading of the fruit that is scarcely less serious to the fruit grower than the injury caused by the presence of mites on the fruit.

[0007] The use of chemicals in miticides is also limited because of their toxicity to

humans, pets, farm animals, non-target vertebrate and invertebrate species, and the environment. Thus, there is a need for miticides effective on plants, shrubs and trees with negligible injury to the foliage and fruit, and with no toxicity to humans, pets, farm animals, non-target vertebrate and invertebrate species, and the environment.

BRIEF SUMMARY OF THE DISCLOSURE

[0008] The present disclosure provides compositions comprising Bulnesia sp. plant

extracts and Humulus sp. plant extracts that can be used as biopesticides, for example, against acari (mite, ticks), nematodes, insects, etc. In some aspects, the compositions disclosed herein comprise a Bulnesia sp. extract and a Humulus sp. plant extract, wherein the Bulnesia sp. extract and the Humulus sp. plant extract act synergistically as miticides.

[0009] In some aspects, the Bulnesia sp. plant is selected from the group consisting of

Bulnesia arborea, Bulnesia bonariensis, Bulnesia carrapo, Bulnesia chilensis, Bulnesia foliosa, Bulnesia loraniensis, Bulnesia macrocarpa, Bulnesia rivas-martinezii, Bulnesia retama, Bulnesia sarmientoi, Bulnesia schickendantzii, and a combination thereof. In some aspects, the Bulnesia sp. plant is Bulnesia sarmientoi. In some aspects, the Humulus sp. plant is selected from the group consisting of Humulus japonicus, Humulus lupulus, Humulus yunnanensis, and a combination thereof. In some aspects, the Humulus sp. plant is Humulus lupulus.

[0010] In some aspects, the Humulus sp. plant extract is obtained from Humulus sp. leaves. In some aspects, the Humulus sp. leaves are fresh leaves or dry leaves. In some aspects, the fresh leaves or dry leaves are, e.g., shredded, fragmented, pulverized, or pelletized. [0011] In some aspects, the Humulus sp. plant extract is obtained from Humulus sp.

flowers or buds (hop cones) or other plant parts (e.g., roots, stems, bark, etc.). In some aspects, the Humulus sp. plant extract is a supercritical C0₂ extract. In some aspects, the Humulus sp. plant extract is obtained by solvent extraction (e.g., using water, organic solvents, or any combination thereof), soxhlet, ultrasonic extraction, or any other method known in the art.

[0012] In some aspects, the Humulus sp. plant extract consists or consists essentially of a blend of hop acids with a high concentration of iso alpha acids. Such blend of hop acids in in some aspects a plant extract that has been enriched in iso alpha acids (e.g., by conversion of alpha acids to iso alpha acids, removal on non-iso alpha acid components, addition of iso alpha acids, or any combination thereof). In some aspects, the

concentration of iso alpha acids is at least 75%. In some aspects, the concentration of iso alpha acids is at least 90%. In some aspects, the iso alpha acids are derived from humulone, adhumulone, cohumulone, posthumulone, prehumulone, or any combination thereof.

[0013] In some aspects, the Bulnesia sp. plant extract is obtained from Bulnesia sp. wood, root, stems, leaves, and other plant parts. In a specific aspect, the Bulnesia sp. plant extract is obtained from Bulnesia sp. wood. In some aspects, the Bulnesia sp. plant extract is obtained by steam flow extraction. In some aspects, the Bulnesia sp. plant extract is obtained by solvent extraction (e.g., using water, organic solvents, or any combination thereof), soxhlet, ultrasonic extraction, or any other method known in the art. In some aspects, the Bulnesia sp. plant extract is an essential oil.

[0014] In some aspects, the Bulnesia sp. plant extract comprises guaiol, bulnesol,

hanamyol, or any combination thereof. In some aspects, the Bulnesia sp. plant extract further comprises a-guaiene, β-caryophyllene, β-guaiene, guaioxide, a-bulnesene, elemol, germacrene-B, eudesm-5-en-l l-ol, B-eudesmol, lO-epi-y-eudesmol, γ-eudesmol, a-eudesmol, 7-epi-a-eudesmol, guaiol, bulnesol, hanalyol. or any combination thereof. In some aspects, the Bulnesia sp. plant extract is an essential oil from Bulnesia sarmientoi wood and the Humulus sp. plant extract is a blend of hop acids extracted from Humulus lupulus leaves with a concentration of iso alpha acids of at least 75%. In some aspects, a formulation for use according to the methods disclosed herein (e.g., as an agricultural, veterinary, or medical acaricide, miticide, insecticide, or nematicide) comprises (i) a Bulnesia sarmientoi plant extract (e.g., an essential oil) comprising a-guaiene, β- caryophyllene, β-guaiene, guaioxide, a-bulnesene, elemol, germacrene-B, eudesm-5-en- 11 -ol, B-eudesmol, lO-epi-y-eudesmol, γ-eudesmol, a-eudesmol, 7-epi-a-eudesmol, guaiol, bulnesol, hanalyol. or any combination thereof, and (ii) a Humulus lupulus plant enriched in iso alpha acids, e.g., a concentration of iso alpha acids of at least 75% expressed as weight/weight (w/w) or weight/volume (w/v).

[0015] In some aspects, the ratio of essential oil from Bulnesia sarmientoi wood and extract from Humulus lupulus leaves enriched in iso alpha acids is about 6: 1 to 1 :6. In some aspects, such ratio is from about 6: 1 to about 4: 1. In some aspects, the ratio of essential oil from Bulnesia sarmientoi wood and extract from Humulus lupulus leaves enriched in iso alpha acids is about 5: 1. In a particular aspect, the ratio of essential oil from Bulnesia sarmientoi wood and extract from Humulus lupulus leaves enriched in iso alpha acids is 5.4: 1.

[0016] In some aspects, the miticide composition further comprises at least a solvent, a surfactant, an emulsifier, a carrier, or a combination thereof. In some aspects, the surfactant or emulsifier is a cationic, anionic or nonionic surfactant or combinations thereof. In some aspects, the surfactant or emulsifier is a polysorbate-type nonionic surfactant (TWEEN®) or sorbitan-type nonanionic surfactant (SPAN®). In some aspects, the surfactant or emulsifier comprises, consists, or consists essentially of TWEEN® 85 (polyoxyethylenesorbitan trioleate). In some aspects, the carrier is selected from the group consisting of canola oil, castor oil, soybean oil, sunflower oil, mineral oil, long chain fatty acid ester, sesame oil, and combination thereof. In some aspects, the long chain fatty acid ester is selected from octanoyl glycerate, methyl oleate, and a

combination thereof. In some aspects, the carrier comprises canola oil. In some aspects, the solvent is selected from the group consisting of ethanol, isopropyl alcohol, butanol, propanol, glycerol, methanol, ethyl lactate, and combinations thereof. In some aspects, the solvent comprises ethanol.

[0017] The present disclosure also provides a method for controlling or preventing mite infestation and/or mite damage to plants, comprising treating the plants or the locus surrounding the plants with a miticidally effective amount of any of the miticide compositions disclosed herein, or a miticide formulation comprising any of the miticide compositions disclosed herein. [0018] Also provided is a method for controlling or preventing insect infestation and/or insect damage to plants, comprising treating the plants or the locus surrounding the plants with an effective amount of any of the miticide compositions disclosed herein, or a formulation comprising of any of the miticide compositions disclosed herein.

[0019] Also provided is a method for controlling or preventing nematode infestation and/or nematode damage to plants, comprising treating the plants or the locus

surrounding the plants with an effective amount of any of the miticide compositions disclosed herein, or a formulation comprising of any of the miticide compositions disclosed herein, adapted for nematicide use.

[0020] Also provided is a method to treat a surface to prevent mite infestation with a miticidally effective amount of any of the miticide compositions disclosed herein, or a formulation comprising of any of the miticide compositions disclosed herein.

[0021] Also provided is a method to treat a surface to prevent insect infestation with an effective amount of any of the miticide compositions disclosed herein, or a formulation comprising of any of the miticide compositions disclosed herein.

[0022] Also provided is a method to treat a surface to prevent nematode infestation with an effective amount of any of the miticide compositions disclosed herein, or a formulation comprising of any of the miticide compositions disclosed herein.

[0023] In some aspects, the mite is Tetranychus urticae. In some aspects, the insect is selected from the group consisting of white flies, trips and aphids. In some aspects, the nematode is selected from the group consisting of Meloidogyne sp., Prtaylenchus sp., and Radopholus sp. In some aspects, the white fly belongs to the genus Trialeurodes, e.g., Trialeurodes vaporariorum. In some aspects, the insect is a leafminer. In some aspects,, the leafminer belongs to the genus Liriomyza, e.g., Liriomyza huidobrensis.

[0024] In some aspects, the surface is in an agricultural, a horticultural, or a garden

environment. In some aspects, the methods disclosed herein further comprise a dose regimen. In some aspects, the dose regimen comprises at least one daily dose. In some aspects, the dose regimen comprises at least one weekly dose. BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES

[0025] FIGS. 1 A-1C show the chemical structures of alpha acids (FIG. 1 A), beta acids

(FIG. IB), and hexehydro-iso alpha acids (FIG. 1C), which are compounds that are present in Humulus sp. plant extracts (e.g., extracts from Humulus lupulus).

[0026] FIGS. 2A-2C show the chemical structures of R- and S- linalool (FIG. 2 A),

xanthohumol (FIG. 2B), and quercetin and cetechine (FIG. 2C), which are compounds that are present in Humulus sp. plant extracts (e.g., extracts from Humulus lupulus).

[0027] FIGS. 3 A and 3B show the chemical structures of 8-prenylaringenin (FIG. 3 A), and mycene, caryophyllene, and humulene (FIG. 3B), which are compounds that are present in Humulus sp. plant extracts (e.g., extracts from Humulus lupulus).

[0028] FIG. 4 shows the chemical structures of guaiol, bulnesol, and hanamyol, which are compounds that are present in extracts from Bulnesia sp. (e.g., Bulnesia sarmientoi plant extracts).

[0029] FIG. 5 shows that formulations comprising a Bulnesia sp. (Bulnesia sarmientoi) plant extract and & Humulus sp. {Humulus lupulus) plant extract function synergistically to control leafminers (Liriomyza huidobrensis).

[0030] FIG. 6 shows that formulations comprising a Bulnesia sp. {Bulnesia sarmientoi) plant extract and & Humulus sp. {Humulus lupulus) plant extract function synergistically to control white flies {Trialeurodes vaporariorum).

[0031] FIG. 7 shows that formulations comprising a Bulnesia sp. {Bulnesia sarmientoi) plant extract and a Humulus sp. {Humulus lupulus) plant extract function synergistically to control nematodes {Meloidogyne sp.).

DETAILED DESCRIPTION

[0032] The present disclosure is directed to miticide compositions comprising a Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) and a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof), wherein the Bulnesia sp. plant extract (obtained, e.g., from Bulnesia sarmientoi) and the Humulus sp. plant extract (obtained, e.g., from Humulus lupulus) function synergistically as miticides. [0033] Also provided are methods of making the miticide compositions disclosed herein, methods of use of such miticide compositions, and articles of manufacture and kits comprising such miticide compositions. In some aspects, the compositions disclosed herein can be used to treat fabrics or different surfaces in order to reduce levels of allergens derived from mites. Furthermore, in addition to agricultural and horticultural uses, the present disclosure provides medical, cosmetic, and veterinary uses for the disclosed compositions (e.g., to treat or prevent mite infections in humans and animals).

[0034] Also provided are uses of compositions comprising a Bulnesia sp. plant extract

(e.g., an extract, or a purified fraction or partially purified fraction thereof) and a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof), for example, as insecticides, fungicides, nematicides, or bactericides.

[0035] In order to provide a clear understanding of the specification and claims, the

following definitions are provided below.

1. Definitions

[0036] As used herein, the singular terms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise. The terms "a" (or "an"), as well as the terms "one or more," and "at least one" can be used interchangeably herein.

[0037] Furthermore, "and/or" where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term "and/or" as used in a phrase such as "A and/or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and/or" as used in a phrase such as "A, B, and/or C" is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0038] As used herein, the term "comprising" means including, made up of, and

composed of. Wherever aspects are described herein with the language "comprising," otherwise analogous aspects described in terms of "consisting of and/or "consisting essentially of are also provided

[0039] All numbers in this description indicating amounts, ratios of materials, physical properties of materials, and/or use are to be understood as modified by the word "about," except as otherwise explicitly indicated. [0040] The term "about" as used in connection with a numerical value throughout the specification and the claims denotes an interval of accuracy, familiar and acceptable to a person skilled in the art. In general, such interval of accuracy is ± 10 %. Thus, "about ten" means 9 to 11.

[0041] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure is related. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press, provide one of skill with a general dictionary of many of the terms used in this disclosure.

[0042] Units, prefixes, and symbols are denoted in their Systeme International de Unites

(SI) accepted form. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, amino acid sequences are written left to right in amino to carboxy orientation. The headings provided herein are not limitations of the various aspects or aspects of the disclosure, which can be had by reference to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by reference to the specification in its entirety.

[0043] The term "plant" as used herein refers to a living organism exemplified by, for example, trees, shrubs, grasses, ferns, flowering plant, vegetables, legumes, or grains. In some aspects, the plant is, for example, a vegetable, a legume, a grain, or a flowering plant. The term plant encompasses any part of a plant, for example fruits, leaves, stems, branches, bark, roots, shoots, seeds, flowers, buds, etc. The term "plant" as used herein encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, shoots, stems, leaves, roots (including tubers), fruits, stalk, seedlings, tubers, flowers, and tissues and organs.

[0044] The terms "Bulnesia sp. " and "Humulus sp." refer to any plant belonging the

genus Bulnesia (e.g., Bulnesia sarmientoi) or to the genus Humulus (e.g., Humulus lupulus), respectively.

[0045] The phrase "Bulnesia sp. plant extract" as used herein refers to a plant extract comprising biologically active substances (e.g., substances with miticide, nematicide, bactericide, fungicide, or insecticide activity) obtained from a plant belonging to the genus Bulnesia, e.g., Bulnesia sarmientoi, using any extraction method known in the art. The term also encompasses purified and/or semi-purified chemical components present in natural Bulnesia sp. extracts, and/or compositions comprising synthetic and/or semisynthetic biologically active substances (e.g., substances with miticide, nematicide, bactericide, fungicide, or insecticide activity) present in natural Bulnesia sp. extracts.

[0046] The phrase "Humulus sp. plant extract" as used herein refers to a plant extract comprising biologically active substances (e.g., substances with miticide, nematicide, bactericide, fungicide, or insecticide activity) obtained from a plant belonging to the genus Humulus, e.g., Humulus lupulus, using any extraction method known in the art. The term also encompasses purified and/or semi-purified chemical components present in natural Humulus sp. extracts, and/or compositions comprising synthetic and/or semisynthetic biologically active substances (e.g., substances with miticide, nematicide, bactericide, fungicide, or insecticide activity) present in natural Humulus sp. extracts.

[0047] The term "plant extract" as used herein, e.g., as in "Bulnesia sp. plant extract" and

"Humulus sp. plant extract," refers to an extract or concentrate that can be obtained from any part of the plant, e.g., seeds, leaves, fruit, stems, bark, roots, or parts thereof {e.g., seed husks, fruit skins, etc.), or combinations thereof, and contains an appreciable amount by weight of the total weight of the extract of a biologically active substance (e.g., a substance with miticide, nematicide, bactericide, fungicide, or insecticide activity), e.g., alpha acids and/or iso alpha acids from Humulus lupulus. In some aspects, the plant extract is, for example, obtained from the leaves. In some aspects, the plant extract is, for example, obtained from wood. In some aspects, the plant extract is obtained from multiple parts of the plant (e.g., leaves and wood). In some aspects, a plants extract is enriched in a biologically active substance (e.g., a substance with miticide, nematicide, bactericide, fungicide, or insecticide activity), e.g., alpha acids and/or iso alpha acids from Humulus lupulus, with respect to the original parts of the plant.

[0048] The enrichment in a biologically active substance (e.g., a substance with miticide, nematicide, bactericide, fungicide, or insecticide activity) can be achieved by a process that increases amounts of one or more components of the plant relative to the percentages that occur naturally in the plant, for example, via extraction {e.g., using solvents) or by concentration. [0049] The method of extraction determines the final composition of the extract.

Numerous methods to extract molecular components from plants tissues are known in the art, e.g., maceration, infusion, digestion, decoction, percolation, repercolation, hot continuous extraction, counter-current extraction, ultra sound extraction (sonication), solvent extraction using water (aqueous extraction), extraction with organic solvents, extraction with mixtures of water and organic solvents, steam distillation, dry distillation, vacuum distillation, hydrodistillation (HD), simultaneous distillation solvent extraction (SDE), microwave assisted hydrodistillation (MWHD), supercritical fluid extraction (SFE), expression ("cold press"), thin layer chromatography (TLC), gel filtration chromatography (GFC), size exclusion chromatography (SEC), flash chromatography, SEPHADEX™ chromatography, high performance liquid chromatography (HPLC), soxhlet extraction, turbo-extraction, carbon-dioxide (C0₂) supercritical

(temperature/pressure) extraction, etc.

[0050] Each one of these methods can produce an extract that will have different physico- chemical characteristics respect to extracts produced by other methods. See Sasidharan et al. (2011), "Extraction, Isolation and Characterization of Bioactive Compounds from Plant's Extracts" Afir. J. Tradit. Complement. Altern. Med. 8(1): 1-10), Azwanida (2015) "A Review on the Extraction Methods Use in Medicinal Plants, Principle, Strength and Limitation." Med. Aromat. Plants 4: 196), and Bart (2011) "Extraction of Natural Products from Plants - An Introduction," in INDUSTRIAL SCALE NATURAL PRODUCTS EXTRACTION (eds H.-J. Bart and S. Pilz), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany).

[0051] Solvents used in the extraction of plant components must be capable of

penetrating the tissues of the plant sample and dissolve the active principles. For example, water alone can extract proteins, glycosides, enzymes, sugars, etc. but will not extract many alkaloids, glycosides, etc. Organic solvents alone can extract gums, waxes, fats, etc. but will not dissolve glycosides, sugars, etc. Accordingly, two extracts generated using the same sample, e.g., leaves, will have a different composition depending on the solvent system used. For example, an extract generated by aqueous extraction (i.e., using only water) inevitably will have a different composition than a extract generated using an organic solvent alone, or than an extract generated by maceration using a water: organic solvent mixture (e.g., water: ethanol). [0052] In specific aspects, the compositions disclosed in the present specification are prepared using extraction methods known in the art that employ organic solvents or pressurized C0₂ (liquid or supercritical). The extracts can then further be evaporated and thus concentrated to yield by means of air drying, vacuum oven drying, fluid-bed drying or freeze-drying, the extract product. Extracts can be further partially purified, e.g. by chromatographic methods. Chemical modification of extracts or partially purified extracts is also possible.

[0053] The Humulus sp. plant extracts and Bulnesia sp. plant extracts disclosed herein can be prepared according to any of these methods and any other methods known in the art. The presence of biologically active substances, e.g. substances with miticide, nematicide, bactericide, fungicide, or insecticide activity, in an extract prepared using any of the methods disclosed above or other methods known in the art which would allow the extract to be used in the compositions and methods disclosed herein, can be determined using methods known in the art (or the methods disclosed in the present application) herein without undue experimentation. For example, whether a Bulnesia sp. or a Humulus sp. plant extract has miticide properties can be determined without undue experimentation according to the methods disclosed in Example 1. The methods presented in Example 1 also allow to determine without undue experimentation whether a combination of Bulnesia sp. or a Humulus sp. plant extracts functions synergically as a miticide (or, for example, as an insecticide, nematicide, bactericide, or fungicide).

[0054] In some aspects, the plant extract is a "juice." The term "juice" as used herein refers to a liquid obtained from the crushing, mashing, pulverizing, grinding, smashing or any combination thereof, plant materials, such as the plant fruit.

[0055] The term "alpha acids" as used herein refers to a class of chemical compounds found mainly in the resin glands of the flowers or buds (hop cones) of the hop plant {Humulus sp.). Alpha acids are the source of hop bitterness. Common alpha acids include humulone, adhumulone, cohumulone, posthumulone, and prehumulone. Alpha acids and iso alpha acids from Humulus sp. are disclosed in further detail below.

[0056] As used herein, the term alpha acids refers not only to alpha acids obtained from natural sources, but also to synthetic alpha acids, semisynthetic alpha acids {e.g., alpha acids chemically and/or enzymatically modified), and combinations thereof. In some aspects, the term alpha acids refers to a single alpha acid species, whereas in other cases (especially when referring to alpha acids from plant extracts), the term alpha acids can refer to a single alpha acid or to a combination of alpha acids.

[0057] The term "iso alpha acids" as used herein refers to a class of chemical compounds derived from alpha acids mainly found in the resin glands of the flowers (hop cones) of hop plants {Humulus sp.). Iso alpha acids are generated by the isomerization of alpha acids present in hops as a result of the application of heat in solution. The degree of isomerization is highly dependent on the length of time the hops are subjected to high temperatures. Longer boil times will result in isomerization of more of the available alpha acids. The most common iso-alpha acids are cis- and trans- isohumulone.

[0058] In some aspects, the compositions disclosed herein comprise Humulus sp. plant extracts enriched iso alpha acids, wherein the iso alpha acid content in the plant extract is at least about 50%, at least about 55%, at least about 60%>, at least about 65%>, at least about 70%), at least about 75%, at least about 80%>, at least about 85%>, at least about 90%, at least about 95%, at least 98%, or at least about 99% of the plant extract expressed as weight/weight (w/w) or weight/volume (w/v).

[0059] The terms "mite" and "mites" as used herein refer in general, to members of

arthropods belonging to the subclass Acari (also known as Acarina). In some aspects, the term mite also includes ticks. Damage to crops is perhaps the most costly economic effect of mites, especially by the spider mites and their relatives (Tetranychoidea), earth mites (Penthaleidae), thread-footed mites (Tarsonemidae) and the gall and rust mites

(Eriophyidae). Some parasitic forms affect humans and other mammals, causing damage by their feeding, and can even be vectors of diseases, such as scrub typhus, rickettsial pox, Lyme disease, Q fever, Colorado tick fever, tularemia, tick-borne relapsing fever, babesiosis, ehrlichiosis and tick-borne meningoencephalitis. A well-known effect of mites on humans is their role as an allergen and the stimulation of asthma in people affected by respiratory disease.

[0060] In some aspects, the terms mite refers to mites responsible for infestations in

plants of commercial value, e.g., Tetranychus urticae, Tetranychus cinnabarinus,

Tetranychus neocaledonicus, Tropilaelaps clarea, Lepidoglyphus sp., Phytoseiulus persimilis, Neoseiulus californicus, Rhizoglyphus tritici, Brivipalpus californicus, Brevipalpus phoenicis, Brevipalpus obovatus, Brevipalpus lewisi, Tenuipalpus dasples, Raoiella indica, Tetranychus cocosi, Spinacus Pagonis, Aceria hibisci, Aceria guerreronis, Aceria chrysophylli, Eriophyes buceras, Aceria pallida, Polyphagotarsonemus latus, Stenotarsonemus spinki, Acarapis woodi, etc.

[0061] Brevipalpus mites are responsible for diseases such as citrus leprosis, orchid fleck, ligustrum ringspot, coffee ringspot, passion fruit green spot, hibiscus green spot, hibiscus chlorotic spot, malvaviscus spot, ivy green spot, schefflera ringspot, clerodendron chlorotic spot, clerodendron green spot, solanum vioaefolium ringspot, and viola chlorotic spot.

[0062] In some aspects, the term mite refers to spider mites. Spider mites are members of the family Tetranychidae, which includes about 1,200 species. They generally live on the undersides of leaves of plants, where they may spin protective silk webs, and they can cause damage by puncturing the plant cells to feed. Spider mites are known to feed on several hundred species of plants. The best known member of the group is Tetranychus urticae, which has a cosmopolitan distribution, and attacks a wide range of plants, including peppers, tomatoes, potatoes, beans, corn, cannabis, and strawberries. Other species which can be important pests of commercial plants include Panonychus ulmi (fruit tree red spider mite) and Panonychus citri (citrus red mite).

[0063] In some aspects, the term mite refers to species belonging to the genera

Neoschizonobi, Sinobryobia, Marainobia, Bryobia, Toronobia, Pseudobryobia,

Strunkobia, Mezranobia, Eremobryobia, Bryobiella, Hemibryobia, Bryocopsis,

Tetranychopsis, Notonychus, Dolichonobia, Monoceronychus, Mesobryobia,

Hystrichonychus, Parapetrobia, Peltanobia, Tauriobia, Aplonobia, Paraplonobia, Beerella, Magdalena, Porcupinychus, Afronobia, Neotrichobia, Schizonobiella, Schizonobia, Dasyobia, Lindquistiella, Edella, Petrobia, Atetranychus, Synonychus, Eurytetranychus, Eurytetranychoides, Eutetranychus, Meyernychus, Aponychus, Paraponychus,

Sinotetranychus, Anatetranychus, Duplanychus, Eonychus, Crotonella, Tenuipalpoides, Tenuipalponychus, Brevinychus, Sonotetranychus, Mixonychus, Evertella, Panonychus, Allonychus, Schizotetranychus, Yunonychus, Yezonychus, Neotetranychus,

Acanthonychus, Mononychellus, Platytetranychus, Eotetranychus, Palmanychus,

Atrichoproctus, Xinella, Oligonychus, Hellenychus, Tetranychus, or Amphi tetranychus.

[0064] Tetranychus urticae (common names include red spider mite and two-spotted spider mite) feeds on ornamental plants such as roses as well as vegetables and food crops such as peppers, tomatoes, potatoes, beans, corn, or strawberries. [0065] Panonichus is a genus of mites of the family Tetranychidae. Panonychus ulmi

(European red mite) feeds on numerous plants species such as grapes (Vitis vinifera, Vitis lambrusca), roses (Rosa canina, Rosa multiflora, Rosa palustris), pear trees (Pyrus baccata, P. communis, P. pyrifolia, P. sargentii), apple trees (Malus domesticd), Prunus sp. (P. americana, P. armeniaca, P. avium, P. cerasius, P. chinensis, P. divaricate, P. domestica, P. dulcis, P. institias, P. padus, P. persica, P. spinose), carrots (Daucus carota), squash (Cucurbuta pepo, Cucurbita maxima), Citrus sp. (C. aurantifolia, C. aurantium, C. grandis), strawberry (Fragaria vesca), beans (Phaseolus), etc.

[0066] Olygonychus is another genus of mites of the family Tetranychidae comprising about 200 species, including O. aceris (maple spider mite), a pest of maples; O.

afrasiaticus (date palm spider mite), a major pest of the date palm in North Africa and the Middle East, also on grasses, including maize; O. biharensis (cassava red mite), a pest of many cultivated fruits and ornamental plants; O. coffeae (tea red spider mite), considered to be the most serious pest of tea, also a pest of coffee, cotton, and jute, and recorded on cashew, African oil palm, rubber, and several fruit crops; O. gossypii (cotton red mite), a pest of cotton, fruits, beans, ornamentals; O. ilicis (southern red mite, coffee red mite), a pest of many woody ornamentals, especially azaleas and camellias; O. indicus (sugarcane red spider mite, sugarcane leaf mite), a pest of banana and grass crops; O. mangiferus (mango spider mite), a pest of many fruit crops, sweet potato, cotton, ornamentals; O. mcgregori, a pest of avocado, cassava, cotton, ornamentals; O. milleri (pine spider mite), a pest of many species of pine; O. oryzae (paddy leaf mite, rice leaf mite), a pest of rice; O. palus, a pest of banana; O. perditus, a pest of ornamental conifers, such as juniper bonsai; O. perseae (persea mite), a top pest of avocado in California, also on ornamentals; O. pratensis (Banks grass mite), a pest of corn, turfgrasses; O. punicae (ash flower gall mite, avocado brown mite), a pest of avocado; O. sacchari (sugarcane yellow mite), a pest of sugarcane and other grasses; O. shinkajii, a pest of grass crops such as rice, sugarcane, and bamboo; O. thelytokus, a pest of some fruits, cassava, cotton, and ornamentals; O. ununguis (spruce spider mite), a pest of conifers with a preference for spruces; O. yothersi (avocado red mite), a pest of yerba mate and avocado; or O. zeae (maize spider mite), a pest of banana and grasses.

[0067] Bryobia is another genus of mites in the Tetranychidae spider mite family.

Perhaps the best known species is Bryobia praetiosa, the clover mite or brown clover mite, an economically important pest of over 250 species of plants. It occurs nearly worldwide. Like other bryobia mites, this species is herbivorous, but there have been occasional reports of it occurring as an ectoparasite on humans, particularly children. It causes an itchy skin irritation. Bryobia artemisiae lives on wormwoods and causes a gray discoloration of the leaves. Bryobia attica feeds on olive. Bryobia convolvulus is commonly found on field bindweed {Convolvulus arvensis), and it is also a pest of alfalfa. Bryobia cristata is reported from nearly every continent. It occurs on many grasses, including wheat and barley. It is also found on azalea, spiderwort, clover, and alfalfa. Bryobia graminum is a pest of fruit trees, clover, and grasses. Bryobia kissophila is a pest of common ivy (Hedera helix) and potato crops. Bryobia lagodechiana may invade greenhouses and it infests beans, cucumbers, currants, and several other cultivated plants. Bryobia rubrioculus damages fruit trees, especially almond and apricot. It is also a pest of vineyards. Bryobia vasiljevi occurs on a variety of crops, including apples, asparagus, and wheat. Bryobia watersi has been found on mallow, cucumber, peanut, and Bermuda grass.

[0068] In some aspects, the term mite includes ticks from the Ixodidae family (hard ticks) or from the Argasidae family (soft ticks). In some aspects, the tick is black-legged or deer tick (Ixodes scapularis), Ixodes pacificus, Ixodes ricinus, or Ixodes persulcatus, which are vectors for Lyme disease. In other aspects, the tick is Ornithodoros sp., which is a vector for relapsing fever. In other aspects, the tick is Dermacentor variabilis, Dermacentor andersoni, or Amblyomma cajennense, which are vectors for Rocky Mountain Spotted Fever. In other aspects, the tick is Amblyomma americanum, a vector for Ehrlichiosis anaplasmosis and Severe Febrile Illness. In other aspects, the tick is Hyalomma marginatum, or Rhipicephalus bursa, which are vectors for Crimean-Congo hemorrhagic fever.

[0069] In some aspects, the term mite refers to members of the superorder Acariformes.

In some aspects, the term mite refers to the members of the order Trombidiformes, which include medically important mites (such as Demodex, the chiggers, and scrub-itch mites) and many agriculturally important species, including the spider mites (Tetranychidae), flat mites (Tenuipalpidae) and gall mites (Eriophyidae).

[0070] In some aspects, the term mite refers to mite that parasite animals such as ear mites (Otodectes cynotis), rabbit ear mites (Psoroptes cuniculi), or scabies (Sarcoptes scabiei). In some aspects, the term mite refers to chiggers (e.g., Trombicula alfreddugesi, Trombicula autumnalis). In some aspects, the term mite refers to the bee parasite Varroa destructor.

[0071] The term "crude" as applied to the plant extract disclosed herein refer to an extract obtained from Humulus sp. or Bulnesia sp. which has not been further processed to increase the concentration of a particular active principle.

[0072] The term "pure" as used herein refers to a composition {e.g., an iso alpha acid- containing Humulus sp. plant extract) enriched at least 60% of active principle {e.g., iso alpha acids) in dry weight. Numerous methods are known in the art to determine purity. See e.g., L. R. Snyder, J.J. Kirkland, and J. W. Dolan, "Introduction to Modern Liquid Chromatography," John Wiley & Sons, New York, (2009) and Rabi, et al, Physical Review 53 (4): 318-327 (1938).

[0073] In some aspects, the term "active principle" refers to a crude Humulus sp. plant extract or a crude Bulnesia sarmientoi plant extract. In other aspects, the term "active principle" refers to one or more biologically active substances (e.g., a substance with miticide, nematicide, bactericide, fungicide, or insecticide activity activity) purified from a crude Humulus sp. plant extract or a crude Bulnesia sarmientoi plant extract. In some aspects, the term active principle refers to a Humulus sp. plant extract prepared according to the method of Example 2 or a variant thereof, & Bulnesia sp. plant extract prepared according to the method of Example 3 or a variant thereof, or a combination thereof.

2. Miticide compositions comprising Bulnesia sp. and Humulus sp. plant extracts

[0074] The present disclosure provides miticide compositions comprising a Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) and a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof), wherein the Bulnesia sp. plant extract (obtained, e.g., from Bulnesia sarmientoi) and the Humulus sp. plant extract (obtained, e.g., from Humulus lupulus) function synergistically as miticides.

[0075] Natural agents can be used as an alternative to pesticides to control mites. The use of synergistic combinations overcomes the need to used higher concentrations of either Bulnesia sp. plant extracts and/ 'or Humulus sp. plant extracts, therefore reducing the risk of toxicity to the plants, other invertebrates, vertebrates, or the environment, and decrease the cost associated with the miticide.

[0076] In some aspects, the Bulnesia sp. plant extract is obtained from Bulnesia arborea

(Maracaibo lignum vitae), Bulnesia bonariensis, Bulnesia carrapo, Bulnesia chilensis, Bulnesia foliosa, Bulnesia loraniensis, Bulnesia macrocarpa, Bulnesia rivas-martinezii, Bulnesia retama, Bulnesia sarmientoi (Argentine lignum vitae, Paraguay lignum vitae, "palo santo", ibiocai), Bulnesia schickendantzii, or a combination thereof. Any part of the plant or combination thereof can be used to produce the Bulnesia sp. plant extract. In a specific aspect, the Bulnesia sp. plant extract is obtained from wood. In a particular aspect, the wood is dry wood.

[0077] In some aspects, the Humulus sp. plant extract is obtained from Humulus

japonicus (Asian Hop), Humulus lupulus (Common Hop), Humulus yunnanensis (Yunnan Hop), or a combination thereof. In some aspects, t e Humulus sp. plant extract comprises a Humulus lupulus extract obtained from Humulus lupulus var. lupulus, Humulus lupulus var. cordifolius, Humulus lupulus var. lupuloides (syn. H. americanus), Humulus lupulus var. neomexicanus, Humulus lupulus var. pubescens, or a combination thereof. Any part of the plant or combination thereof can be used to produce the Humulus sp. plant extract. In a specific aspect, the Humulus sp. plant extract is obtained from leaves. In a particular aspect, the leaves are fresh leaves.

[0078] In some aspects, the Bulnesia sp. plant extract can be obtained from a plant, and can be used in extract, concentrate, or purified form. In some aspects, the Bulnesia sp. plant extract comprises, consists, or consists essentially of (i) miticide compounds present in Bulnesia sp. that have been obtained from the Bulnesia sp. plant (or derivatives thereof), (ii) miticide compounds present in Bulnesia sp. that have been chemically synthesized, or (iii) combinations thereof.

[0079] Similarly, in some aspects, the Humulus sp. plant extract can be obtained from a plant, and can be used in extract, concentrate, or purified form. In some aspects, the Humulus sp. plant extract comprises, consists, or consists essentially of (i) miticide compounds present in Humulus sp. that have been obtained from the Humulus sp. plant (or derivatives thereof), (ii) miticide compounds present in Humulus sp. that have been chemically synthesized, or (iii) combinations thereof. [0080] In some aspects, the miticide compositions disclosed herein can exert their action by contact, ingestion, or both. In some aspects, the miticide compositions disclosed herein are applied at concentrations that are no phytotoxic. In some aspects, the miticide compositions disclosed herein are highly stable, with a shelf life of at least lyear or at least 2 years, and a field residuality of at least 1 week, 2 weeks, 3 weeks or 4 weeks under dry conditions.

[0081] The main chemical components in the Bulnesia sp. plant extracts disclosed herein are guaiol, bulnesol, and hanamyol. Other components of the Bulnesia sp. plant extracts disclosed herein are, for example, a-guaiene, β-caryophyllene, β-guaiene, guaioxide, a- bulnesene, elemol, germacrene-B, eudesm-5-en-l l-ol, B-eudesmol, lO-epi-y-eudesmol, γ- eudesmol, a-eudesmol, and 7-epi-a-eudesmol. The Bulnesia sp. plant extract components in the miticide compositions disclosed herein can be natural, synthetic, semisynthetic, or a combination thereof.

[0082] Similarly, the main active components in the Humulus sp. plant extracts disclosed herein are iso alpha acids. Iso alpha acids are degradation product of naturally occurring alpha acids, which degrade when subjected to high temperature. For example, as show in the reaction below, humulone (a-lupulic acid) can be degraded to the cis- and trans- isohumulone iso alpha acids.

[0083] Thus, the treatment of the crude Humulus sp. plant extracts with high temperature results in marked structural differences from the corresponding natural products. For example, alpha acids are structurally transformed into iso alpha acids, resulting in a higher concentration of iso alpha Humulus sp. plant extracts with respect to the concentration present in crude Humulus sp. plant extracts. Also, iso alpha acids (e.g., cis- and trans-isohumulone iso alpha acids) are significantly more soluble than their corresponding naturally occurring alpha acid (humulone).

[0084] The Humulus sp. plant extract components in the miticide compositions disclosed herein can be natural, synthetic, semisynthetic, or a combination thereof. As used herein the term semisynthetic refers to a molecule that is the result of a chemical or enzymatic modification of a molecule obtained from a natural source (e.g., an alpha acid obtained from a Humulus sp. plant extract can be subsequently modified, either chemically or enzymatically to yield a semisynthetic alpha acid or iso alpha acid).

[0085] In particular, the iso alpha acids in the miticide compositions disclosed herein can be natural (e.g., cis- and trans-isohumulone), synthetic, semisynthetic, or a combination thereof. For example, humulone can be synthesized by the acylation of 1,2,3,5- benzenetetrol with isovaleryl chloride to give 2,3,4,6,-tetrahydroxyisovalerophenone. This step is followed by prenylation with l-bromo-3-methyl-2-butene to give humulone.

[0086] Components of the Humulus sp. plant extracts disclosed herein comprise, for example:

(i) alpha acids such as cohumulone, humulone, or adhumulone, their

respective iso alpha acids, or combinations thereof;

(ii) beta acids such as colupulone, lupulone, or adlupulone, their respective iso beta acids, or combinations thereof;

(iii) hexahydro-isoalpha acids such as isopropyl, isobutyl, or sec-butyl

hexahydro-isoalpha acids;

(iv) R-linalool, S-linalool, canthohumol, quercetin, catechine, 8- prenylnaringenin, myrcene, humulene, caryophyllene; or,

(v) combinations thereof.

[0087] In some aspects, the miticide compositions disclosed herein comprises

(i) a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract); and,

(ii) a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract)

enriched in iso alpha acids (e.g., comprising at least about 60% iso alpha acids w/w or w/v),

wherein the Bulnesia sp. plant extract and the Humulus sp. plant extract act synergistically as miticides.

[0088] In some aspects, the Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) consists or consisting essentially of Humulus sp. purified iso alpha acids.

[0089] In some aspects, a pure composition comprises at least about 60%>, at least about

65%), at least about 70%, at least about 75%, at least about 80%>, at least about 85%>, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% of active principle (e.g., iso alpha acids) expressed as w/w or w/v.

[0090] In some aspects, the miticide composition comprises a Humulus sp. plant extract

(e.g., a Humulus lupulus plant extract), and a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract), with a concentration of Humulus sp. plant extract ranging from 0 to about 1.5 g/L, and a concentration of Bulnesia sp. plant extract ranging from 0 to about 5 g/L. In some aspects, the Humulus sp. plant extract is prepared according to the method of Example 2. In some aspects, the Bulnesia sp. plant extract is prepared according to the method of Example 3. In some aspects, the Bulnesia sp. plant extract and the Humulus sp. plant extract act synergistically as miticides.

[0091] In some aspects, the Humulus sp. plant extract (e.g., a Humulus lupulus plant extract)concentration in the composition is at least about 0.1 g/L, at least about 0.2 g/L, at least about 0.3 g/L, at least about 0.4 g/L, at least about 0.5 g/L, at least about 0.6 g/L, at least about 0.7 g/L, at least about 0.8 g/L, at least about 0.9 g/L, at least about 1 g/L, at least about 1.1 g/L, at least about 1.2 g/L, at least about 1.3 g/L, at least about 1.4 g/L, at least about 1.5 g/L, at least about 1.6 g/L, at least about 1.7 g/L, at least about 1.8 g/L, at least about 1.9 g/L, at least about 2 g/L, at least about 2.1 g/L, at least about 2.2 g/L, at least about 2.3 g/L, at least about 2.4 g/L, or at least about 2.5 g/L. In some aspects, the Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) is an extract prepared according to the method of Example 2.

[0092] In some aspects, the Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) concentration is at least about 0.1 g/L, at least about 0.2 g/L, at least about 0.3 g L, at least about 0

g L, at least about 0

g/L, at least about 1

g/L, at least about 2

g/L, at least about 3

g/L, at least about 4 g/L, at least about 4.4 g/L, at least about 4.5 g/L, at least about 4.6 g/L, at least about 4.7 g/L, at least about 4.8 g/L, at least about 4.9 g/L, at least about 5 g/L, at least about 5.1 g/L, at least about 5.2 g/L, at least about 5.3 g/L, at least about 5.4 g/L, at least about 5.5 g/L, at least about 5.6 g/L, at least about 5.7 g/L, at least about 5.8 g/L, at least about 5.9 g/L, at least about 6 g/L, at least about 6.1 g/L, at least about 6.2 g/L, at least about 6.3 g/L, at least about 6.4 g/L, at least about 6.5 g/L, at least about 6.6 g/L, at least about 6.7 g/L, at least about 6.8 g/L, at least about 6.9 g/L, at least about 7 g/L, at least about 7.1 g/L, at least about 7.2 g/L, at least about 7.3 g/L, at least about 7.4 g/L, or at least about 7.5 g/L. In some aspects, the Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) is an extract prepared according to the method of Example 3.

[0093] In some aspects, the miticide composition disclosed herein comprises a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) wherein the Humulus sp. plant extract at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 1 1%>, at least about 12%>, at least about 13%>, at least about 14%), or at least about 15%> of the composition (w/w or w/v). In some aspects, the Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) is an extract prepared according to the method of Example 2.

[0094] In some aspects, the miticide composition disclosed herein comprises a, Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) wherein the Bulnesia sp. plant extract at least about 10%>, at least about 1 1%>, at least about 12%>, at least about 13%>, at least about 14%>, at least about 15%>, at least about 16%>, at least about 17%>, at least about 18%), at least about 19%>, at least about 20%>, at least about 21%>, at least about 22%, at least about 23%>, at least about 24%>, at least about 25%>, at least about 26%>, at least about 27%o, at least about 28%>, at least about 29%>, at least about 30%>, at least about 3 1%>, at least about 32%>, at least about 33%>, at least about 34%>, at least about 35%>, at least about 36%o, at least about 37%>, at least about 38%>, at least about 39%>, at least about 40%>, at least about 41%>, at least about 42%>, at least about 43%>, at least about 44%>, at least about 45%o, at least about 46%>, at least about 47%>, at least about 48%>, at least about 49%>, or at least about 50%> of the composition (w/w or w/v). In some aspects, the Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) is an extract prepared according to the method of Example 3. [0095] In some aspects, the miticide composition comprises

(A) a Humulus sp. plant extract (e.g. , a plant extract from Humulus lupulus), and

(B) a Bulnesia sp. plant extract (e.g., a plant extract from Bulnesia sarmientoi) in a weight ratio of (A):(B), for example, between 100: 1 to 1 : 100, between 100:1 to 1:75, between 100:1 to 1:50, between 100:1 to 1:25, between 100:1 to 1:10, between 100:1 to 1:5, between 100:1 to 1:1, between 75:1 to 1:75, between 75:1 to 1:50, between 75:1 to 1:25, between 75:1 to 1:10, between 75:1 to 1:5, between 75:1 to 1:1, between, 50:1 to 1:50, between 50:1 to 1:10, between 50:1 to 1:5, between 50:1 to 1:1, between 15:1 to 1:15, between 15:1 to 1:10, between 15:1 to 1:5, between 15:1 to 1:1, between 10:1 to 1:10, between 10:1 to 1:5, between 10:1 to 1:1, or between 6:1 to 1.1, wherein the Humulus sp. plant extract (e.g., a plant extract from Humulus lupulus), and the Bulnesia sp. plant extract (e.g., a plant extract from Bulnesia sarmientoi) act synergistically as miticides.

[0096] In some aspects, the miticide composition disclosed herein comprises

(i) a Bulnesia sarmientoi plant extract, wherein said plant extract is an

essential oil (e.g., prepared according to the method of Example 3), and

(ii) a Humulus lupulus plant extract (e.g., prepared according to the method of Example 2), wherein said plant extract is enriched in iso alpha acids, wherein both plant extracts function synergistically as miticides, and wherein the Bulnesia sarmientoi plant extract and the Humulus lupulus plant extract are at a ratio from about 10:1 (w/w, w/v, v/w, or v/v/) to about 3:1 (w/w, w/v, v/w, or v/v/). In a particular aspect, the ratio is from about 6:1 or about 4:1, e.g., 6:1, 5.9:1, 5.8:1, 5.7:1, 5.6:1, 5.5:1, 5.4:1, 5.3:1, 5.2:1, 5.1:1, 5:1, 4.9:1, 4.8:1, 4.7:1, 4.6:1, 4.5:1, 4.4:1, 4.3:1, 4.2:1, 4.1:1, or 4:1.

3. Methods of making miticide compositions comprising Bulnesia sp. and Humulus sp. plant extracts

[0097] The instant disclosure also provides methods of making miticide compositions comprising a Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) and a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof), wherein the Bulnesia sp. plant extract (obtained, e.g., from Bulnesia sarmientoi) and the Humulus sp. plant extract (obtained, e.g., from Humulus lupulus) function synergistically as miticides. Such methods comprise combining a Bulnesia sp. plant extract and a Humulus sp. plant extract in such a ratio that the observed miticide effect is synergistic. The synergistic nature of the mitidice effect can be determined using the methods disclosed in the instant disclosure (e.g., using Tetranychus mites as a model system) without undue experimentation.

[0098] The manufacture of the disclosed miticide compositions comprises both (i) the production of a miticide composition by combining a Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) and a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof), and (ii) production of a miticide composition by providing separately a Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) and a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) as a kit or article of manufacture together with a set of instructions or packaging indicating how to combine both extracts to produce a miticide composition disclosed herein.

[0099] The Bulnesia sp. plant extracts and/or Humulus sp. plant extracts used to

manufacture the miticide compositions disclosed herein can be produced using any methods know in the art. The Bulnesia sp. plant extracts and/or Humulus sp. plant extracts using to manufacture the miticide compositions disclosed herein can also be obtained from commercial sources.

[0100] In some aspects, the Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) and/or the Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) can be obtained by using extraction methods known in the art {e.g., liquid extraction) using a combination of water and, for example, at least one, at least two, at least three, at least four organic solvents. In some aspects, the Humulus sp. plant extract and/or the Bulnesia sp. plant extract can be obtained by using one extraction method. In some aspects, the Humulus sp. plant extract and/or the Bulnesia sp. plant extract can be obtained by using sequentially more than one extraction method. In some aspects, the sequential extraction methods are all of the same extraction method, e.g., the plant material(s) can be extracted with water in combination with one or more organic solvents {e.g., ethanol or methanol), and after removing the solvent, the remaining plant material(s) can be re-extracted with the same solvent mixture. In other aspects, the Humulus sp. plant extract and/or the Bulnesia sp. plant extract can be obtained using sequentially different extraction methods. In some aspects, the Humulus sp. plant extract and/or the Bulnesia sp. plant extract can be obtained using at least one extraction method that is different than all of the other extraction methods.

[0101] In some aspects, the Humulus sp. plant extract and/or the Bulnesia sp. plant

extract can be obtained, for example, using a liquid-liquid extraction method. A liquid- liquid extraction method is the extraction of a substance from one liquid phase to another liquid phase. In some aspects, the liquid-liquid extraction method comprises, for example, water and an organic solvent. In some aspects, the organic solvent is, for example, methanol, ethanol, isopropanol, methyl acetate, ethyl acetate, isopropyl acetate, n-butyl acetate, isobutyl acetate, acetone, methyl ethyl ketone, methyl isobutyl ketone, dibutyl ether, anisole, toluene, heptane, supercritical C0₂, acetic acid, acetonitrile, benzene, 1- butanol, 2-butanol, 3-butanone, t-butyl alcohol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, 1,2-dichloroethane, diethyl ether, di ethylene glycol, dimethylether, dioxane, ethylene glycol, glycerin, hexane, methyl t-butyl ether, pentane, tri ethyl amine, o-xylene, w-xylene, ^-xylene, cyclopentane, 1,4-dioxane,

dichloromethane, tetrahydrofuran, dimethylformamide, dimethyl sulfoxide, propylene carbonate, formic acid, «-butanol, isopropanol, «-propanol, nitromethane, or any combination thereof.

[0102] In some specific aspects, the Humulus sp. plant extract for use in the miticide compositions disclosed herein is a Humulus lupulus plant extract obtained by subjecting, e.g., Humulus lupulus leaves (for example fresh leaves), to supercritical C0₂ extraction, wherein the content of iso alpha acids in the resulting Humulus sp. plant extract is at least 75%.

[0103] In some specific aspects, the Bulnesia sp. plants extract for use in the miticide compositions disclosed herein is a Bulnesia sarmientoi plan extract obtained by subjecting, e.g., Bulnesia sarmientoi wood to steam flow extraction, wherein the resulting Bulnesia sp. plant extract is an essential oil. The term "essential oil" as used herein refers to a volatile plant oil, non-volatile plant oil, or combination thereof, or a plant extract that is enriched in such components which is water-immiscible. 4. Hamulus sp. plant extracts and Humulus sp. derivatives

[0104] The Humulus sp. plant extracts (e.g., Humulus lupulus plant extracts) used in the compositions disclosed herein comprise alpha acids, and/or iso alpha acids, and/or beta acids. These compounds either occur naturally in Humulus sp. plant, are chemically derived (either through natural biosynthetic processes (e.g., living organism metabolism, e.g., mammal, plant, bacteria), or are derived by synthetic processes using human intervention (e.g., chemical synthesis). Humulus sp. plants contain two major organic acid classes, alpha acids (also known as humulones) and beta acids (also known as lupulones), which contain at least one dissociative moiety (e.g. enolic OH or phenolic OH).

[0105] Alpha acids are organic acids derived from Humulus sp. having structural

homology to a humulone, adhumulone, cohumulone, or an analog or derivative thereof. Humulone, adhumulone, and cohumulone are the three most abundant alpha acid analogs. Other exemplary derivatives of an alpha acid include, but are not limited to isoalpha acids, rhoisoalpha acids, tetrahydroisoalpha acids, and hexahydroisoalpha acids.

[0106] By "beta acid" is meant an organic acid derived from Humulus sp. (e.g., Humulus lupulus) having structural homology to a lupulone, adlupulone, colupulone or an analog or derivative thereof. Lupulone, adlupulone, and colupulone are the three most abundant beta acid analogs. Other exemplary derivatives of a beta acid include, but are not limited to, hulupones, hexahydrobeta acids and hexahydro hulupones. The percentages of analogs present in alpha acids and beta acids is plant variety dependent.

[0107] There are three major analogs for alpha acids (I), humulone, cohumulone, and adhumulone, and three major analogs for beta acids (II), colupulone, lupulone, and adlupulone, as well as the pre- and post-analogs:

ALPHA-ACID R BETA-ACID

Humulone (a) CH₂CH(CH₃)₂ Lupulone (a)

Cohumulone (b) CH(CH₃)₂ Colupulone (b)

Adhumulone (c) CH(CH₃)CH₂CH: Adlupulone (c) Prehumulone (d) CH₂CH₂CH(CH₃)2 Prelupulone (d)

Posthumulone (e) C₂H₅ Postlupulone (e)

[0108] If not otherwise stated, the above meanings for R apply throughout this

specification and also denominate the structure of respective co-, ad-, pre-, and post- derivatives of the compounds listed below, even if such a denomination may not be common in the literature.

[0109] Exemplary Humulus sp. derivatives include iso alpha acids (III), which occur in the cis- and transform (referring to the position of the tertiary alcohol in 4-position of the ring and the renyl side chain in 5-position.

(Ill) cis iso alpha acid (a) trans iso alpha acid (b)

[0110] The resulting compounds are thus cis-iso-humulone, trans-iso-humulone, cis-iso- cohumulone, trans-iso-cohumulone, cis-iso-adhumulone, trans-iso-adhumulone, cis-iso- prehumulone, trans-iso-prehumulone, cis-iso-posthumulone, and trans-iso-posthumulone.

[0111] Humulus sp. derivatives derived from iso-alpha acids are rho-iso-alpha acids (IV)

(also called dihydro iso alpha acids) obtainable by sodium borohydride reduction of iso- alpha acids, tetrahydro-iso-alpha acids (V), obtainable by hydrogenation of iso-alpha acids, and hexahydro iso alpha acids (VI), obtainable by hydrogenation and sodium borohydride reduction of iso-alpha acids.

(IV) (V) (VI) [0112] In view of the chiral center created by the sodium borohydride reduction in the 4- side chain, the compounds are thus: (R)-cis-dihydro-isohumulone, (S)-cis-dihydro- isohumulone, (R)-trans-dihydro-isohumulone, (S)-trans-dihydro-isohumulone, (R)-cis- dihydro-isocohumulone, (S)-cis-dihydro- isocohumulone, (R)-trans-dihydro- isocohumulone, (S)-trans-dihydro-isocohumulone, (R)-cis-dihydro-isoadhumulone, (S)- cis-dihydro-isoadhumulone, (R)-trans-dihydro- isoadhumulone, (S)-trans-dihydro- isoadhumulone; cis-tetrahydro-isohumulone, trans-tetrahydro-isohumulone, cis- tetrahydro-isocohumulone, trans-tetrahydro-isocohumulone, cis-tetrahydro- isoadhumulone, trans-tetrahydro- isoadhumulone; (R)-cis-hexahydro-isohumulone, (S)- cis-hexahydro-isohumulone, (R)-trans-hexahydro- isohumulone, (S)-trans-hexahydro- isohumulone, (R)-cis-hexahydro-isocohumulone, (S)- cis-hexahydro-isocohumulone, (R)- trans-hexahydro-isocohumulone, (S)-trans-hexahydro- isocohumulone, (R)-cis- hexahydro-isoadhumulone, (S)-cis-hexahydro-isoadhumulone, (R)- trans-hexahydro- isoadhumulone, and (S)-trans-hexahydro-isoadhumulone, as well as the respective pre- and posthumulone compounds, preadhumulone (R=C₅Hii), and the compound where R=- CH₂-CH₂-CH(CHs)-CH₂-CH₃, with their respective iso-, dihydro-iso-, tetrahydro-iso-, and hexahydro-iso-derivatives in all isomeric forms.

[0113] Additional Humulus sp. derivatives based on alpha acids are reduced derivatives of alpha acids, such as dihydrohumulone, dihydrocohumulone, dihydroadhumulone, dihydroprehu- mulone, dihydroposthumulone, tetrahydrohumulone,

tetrahydrocohumulone, tetrahydroadhumulone, tetryhydroprehumulone,

tetrahydroposthumulone, humulohydroquinone, cohumulohydroquinone,

adhumulohydroquinone, prehumulohydroquinone, posthumulohydroquinone,

humuloquinone, cohumuloquinone, adhumuloquinone, prehumuloquinone,

posthumuloquinone, 2,6-bis(3-methylbutyl)-4-(COR)resorcinol, 2,6-bis(3-methylbutyl)- 4-(CH₂R)resorcinol; oxidized derivatives of alpha acids, such as humulinone (sometimes termed gamma-acid), cohumulinone, adhumulinone, prehumulinone, posthumulinone, tricyclodehydro-isohumulone, tricyclodedydro-isocohumulone,

tricyclodehydroisoadhumulone, tricyclodehydroisoprehumulone,

tricyclodehydroisoposthumulone; iso alpha acids, such as the compounds (III) described above as well as spiro-isohumulones, spiro-isocohumulones, spiro-isoadhumulones, spiro-isoprehumulones, spiro-isoposthumulones; reduced derivatives of iso-alpha acids, such as compounds (IV), (V), and (VI) described above, as well as neohydro- isohumulones, neohydro-isocohumulones, neohydro-isoadhumulones,

neohydroisoprehumulones, neohydro-isoposthumulones; oxidized derivatives of iso-alpha acids, such as abeo-isohumulones, abeo-isocohumulones, abeo-isoadhumulones, abeo- isoprehumulones, abeo-isoposthumulones; further derivatives of iso-alpha acids, such as allo-isohumulones, allo-isocohumulones, allo-isoadhumulones, allo-isoprehumulones, allo-isoposthumulones, hydrated allo-isohumulones, hydrated allo-isocohumulones, hydrated allo-isoadhumulones, hydrated allo-isoprehumulones, hydrated allo- isoposthumulones, acetylhumulinic acid, acetylcohumulinic acid, acetyladhumulinic acid, acetylprehumulinic acid, acetylposthumulinic acid, humulinic acid (VII), cohumulinic acid, adhumulinic acid, prehumulinic acid, posthumulinic acid, humulinic acid C, cohumulinic acid C, adhumulinic acid C, prehumulinic acid C, posthumulinic acid C, deoxohumulinic acid C, deoxocohumulinic acid C, deoxoadhumulinic acid C,

deoxoprehumulinic acid C, deoxoposthumulinic acid C, humulinic acid D, cohumulinic acid D, adhumulinic acid D, prehumulinic acid D, posthumulinic acid D, O- methylhumulinic acid D, O-methylcohumulinic acid D, O-methyladhumulinic acid D, O- methylprehumulinic acid D, O-methylposthumulinic acid D, O-methylallohumulinic acid D, O-methylallocohumulinic acid D, O-methylalloadhumulinic acid D, O- methylalloprehumulinic acid D, O-methylalloposthumulinic acid D, O- methyldihydrohumulinic acid D, O-methyldihydrocohumulinic acid D, O- methyldihydroadhumulinic acid D, O-methyldihydroprehumulinic acid D, O- methyldihydroposthumulinic acid D, isohumulinic acid, isocohumulinic acid,

isoadhumulinic acid, isoprehumulinic acid, isoposthumulinic acid; reduced derivatives of humulinic acids, such as dihydrohumulinic acid, dihydrocohumulinic acid,

dihydroadhumulinic acid, dihydroprehumulinic acid, dihydroposthumulinic acid, dehydrated dihydrohumulinic acid, dehydrated dihydrocohumulinic acid, dehydrated dihydroadhumulinic acid, dehydrated dihydro- prehumulinic acid, dehydrated

dihydroposthumulinic acid, dehydrated dihydrodeoxohumulinic acid, dehydrated dihydrodeoxocohumulinic acid, dehydrated dihydrodeoxoadhumulinic acid, dehydrated dihydrodeoxoprehumulinic acid, dehydrated dihydrodeoxoposthumulinic acid, dihydrodeoxohumulinic acid, dihydrodeoxocohumulinic acid, dihydrodeoxoadhumulinic acid, dihydrodeoxoprehumulinic acid, dihydrodeoxoposthumulinic acid, deoxoisohumulinic acid, deoxo-isocohumulinic acid, deoxoisoadhumulinic acid, deoxoisoprehumulinic acid, deoxoisoposthumulinic acid; oxidized derivatives of the humulinic acids, such as oxyhumulinic acid, oxycohumulinic acid, oxyadhumulinic acid, oxyprehumulinic acid, oxyposthumulinic acid, dehydrohumulinic acid,

dehydrocohumulinic acid, dehydroadhumulinic acid, dehydroprehumulinic acid, dehydroposthumulinic acid, cyclized dehydrohumulinic acid, cyclized

dehydrocohumulinic acid, cyclized dehydroadhumulinic acid, cyclized

dehydroprehumulinic acid, cyclized dehydroposthumulinic acid; dehydrated humulinic acid, dehydrated cohumulinic acid, dehydrated adhumulinic acid, dehydrated

prehumulinic acid, and dehydrated posthumulinic acid, all of the above compounds, if applicable, as the respective cis (syn) and trans (anti) isomers.

R=CH₂-CH(CH₃) (VII)

[0114] The miticide compositions disclosed herein can also comprise salts of alpha acids and their derivatives, such as salts derived from alkali metals and alkaline earth metals, e.g. sodium, potassium and magnesium salts.

[0115] All of the above compounds are known from the literature and described, e.g. in

D. E. Briggs et al., Brewing: Science and Practice, Woodhead Publishing Ltd., 2004, and M. Verzele and D. De Keukeleire, Chemistry and Analysis of Hop and Beer Bitter Acids, Elsevier, Amsterdam, 1991.

[0116] Further exemplary Humulus sp. derivatives include beta acids, preferably

lupulone, colupulone, adlupulone, prelupulone, and postlupulone, and derivatives of beta acids, including reduced derivatives such as tetrahydrodeoxyhumulone,

tetryhydrodeoxycohumulone, tetrahydrodeoxyadhumulone,

tetrahydrodeoxyprehumulone, tetrahydrodeoxyposthumulone, deoxyhumulone, deoxycohumulone, deoxyadhumulone, deoxyprehumulone, deoxyposthumulone, hexahydrohulupone, hexahydrocohulupone, hexahydroadhulupone,

hexahydroprehulupone, and hexahydroposthulupone, oxidized derivatives of beta acids, such as hulupone (sometimes termed delta-acid), cohulupone, adhulupone, prehulupone, posthulupone, dihydrohulupone, dihydrocohulupone, dihydroadhulupone, dihydroprehulupone, dihydroposthulupone, tetrahydrohulupone, tetrahydrocohulupone, tetrahydroadhulupone, tetrahydroprehulupone, tetrahydroposthulupone,

hexahydrohulupone, hexahydrocohulupone, hexahydroadhulupone,

hexahydroprehulupone, hexahydroposthulupone, hulupinic acid, cohulupinic acid, adhulupinic acid, prehulupinic acid, posthulupinic acid, lupoxes, colupoxes, adlupoxes, prelupoxes, postlupoxes, lupdoxes, colupdoxes, adlupdoxes, prelupdoxes, postlupdoxes, lupdeps, colupdeps, adlupdeps, prelupdeps, postlupdeps, lupdols, colupdols, adlupdols, prelupdols, postlupdols and tricyclo-oxycolupulones (TCOC).

[0117] The miticide compositions disclosed herein can also comprise salts of beta acids and derivatives thereof, such as salts derived from alkali metals or alkaline earth metal salts, e.g. potassium and magnesium salts.

[0118] Alpha acids and beta acids can be prepared by purification from natural hops (e.g.,

Humulus lupulus) and also by chemical synthesis according to traditional methods. The compounds delineated herein, in particular derivatives of alpha and beta acids, can be synthesized using conventional methods known in the art.

[0119] In an aspect of the present disclosure, the composition comprising a Humulus sp. derivative is a Humulus sp. extract, a partially purified Humulus sp. extract (i.e., an extract enriched in one or more components, e.g., iso alpha acids), a chemically modified Humulus sp. extract, or a combination thereof.

[0120] Compositions comprising Humulus sp. derivatives are also available

commercially. John I. Haas, Inc. products containing hops-derivatives include

BETACIDE^®, REDIHOP^®, ISOHOP^®, ISOSTAB^®, TETRAHOP GOLD^®, HEXAHOP GOLD^®, HEXAHOP 95, MGRIAA and MGBETA. The active ingredients in these products are beta acids and alpha acids, rhoisoalpha acids (RIAA, REDIHOP®), isoalpha acids (IAA), tetrahydroisoalpha acids (THIAA), hexahydroisoalpha acids (HHIAA) and tetrahydroisoalpha acids (THIAA) in a 50:50 blend, hexahydroisoalpha acids (HHIAA) and tetrahydroisoalpha acids (THIAA) in a 95:5 blend, magnesium salts of rhoisoalpha acids (MGRIAA) and magnesium salts of beta acids MGBETA), respectively.

[0121] While Humulus sp. plant extracts in the miticide compositions disclosed herein generally contain more than one Humulus sp. derivative (e.g., one or more iso alpha acids), the term derivative as used herein also comprises substantially pure derivatives obtained, e.g., by chemical synthesis or isolation/purification ΐτονα Humulus sp. plant extracts.

[0122] Numerous methods are available for the chemical synthesis of Humulus sp.

derivatives. Such compounds can be synthesized from readily available starting materials using standard synthetic techniques and methodologies known to those of ordinary skill in the art. Synthetic chemistry transformations and protecting group methodologies

(protection and de- protection) useful in synthesizing the compounds identified by the methods described herein are known in the art and include, for example, those such as described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2^nd ed., John Wiley and Sons (1991 ); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), and subsequent editions thereof.

[0123] Chemically synthesized alpha and beta acids can be separated from a reaction mixture and further purified by a method such as column chromatography, high-pressure liquid chromatography, or recrystallization. As can be appreciated by the skilled artisan, further methods of synthesizing the compounds herein will be evident to those of ordinary skill in the art. Additionally, the various synthetic steps may be performed in an alternate sequence or order to give the desired compounds.

[0124] Humulus sp. derivatives used in the compositions disclosed herein can contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, single enantiomers, individual diastereomers and diastereomeric mixtures. All such isomeric forms of these compounds are expressly included in the present disclosure. The Humulus sp. derivatives of the present disclosure can also be represented in multiple tautomeric forms. In such instances, the present disclosure expressly includes all tautomeric forms of the compounds described herein. All such isomeric forms of such Humulus sp. derivatives are expressly included in the present invention. All crystal forms of the Humulus sp. derivatives described herein are expressly included in the present disclosure. As used herein, the Humulus sp. derivatives of the present disclosure are defined to include modifications of naturally occurring Humulus sp. derivatives. [0125] Modifications include Humulus sp. compounds disclosed herein that are modified by appending appropriate functionalities to enhance desired properties. The Humulus sp. derivatives of this disclosure may be modified by appending appropriate functionalities to enhance selective biological properties. Such modifications are known in the art and include those, which increase an anti-parasitic activity (e.g., miticide), increase solubility, or increase heat, light, or oxidation stability. Preferred modifications include salts, esters and amides of naturally occurring Humulus sp. derivatives.

[0126] Acceptable salts of the Humulus sp. compounds disclosed herein include those derived from acceptable inorganic and organic acids and bases. Examples of suitable acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, di gluconate, dodecyl sulfate,

ethanesulfonate, formate, fumarate, glucoheptanoate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other acids, such as oxalic acid, may be employed in the preparation of salts useful as intermediates in obtaining the Humulus sp. compounds disclosed herein.

[0127] In some aspects, the compositions disclosed herein include Humulus sp. acids salts (e.g., sodium, potassium, lithium, calcium, magnesium) having increased stability. These Humulus sp. acid salt (e.g., sodium, potassium, lithium, calcium, magnesium) compositions are advantageously stable relative to Humulus sp. acids produced by conventional methods, which are susceptible to degradation due to heat, light, and acid catalysis.

[0128] Salts derived from appropriate bases include any monovalent cation, including but not limited to lithium, sodium, potassium, silver, copper, or divalent cation, including but not limited to magnesium, calcium, barium, chromium, manganese, iron, cobalt, nickel, copper, zinc, and cadmium. In particular aspects, the salt is derived from alkali metals or alkaline earth metals (e.g., sodium, potassium, lithium, magnesium, calcium, etc.), ammonium and N-(alkyl)₄ salts. Any basic nitrogen-containing groups of the compounds disclosed herein can be quatemizated. Water or oil-soluble or dispersible products may be obtained by such quaternization. [0129] In some aspects, the compositions disclosed herein comprise one or more

Humulus sp. acid derivatives, e.g., free acids or salts thereof.

[0130] In one aspects, the compositions disclosed herein comprise alpha acids, reduced derivatives of alpha acids, oxidized derivatives of alpha acids, iso alpha acids, reduced derivatives of iso alpha acids, oxidized derivatives of iso alpha acids, humulinic acids, isomers of humulinic acids, reduced derivatives of humulinic acids, oxidized derivatives of humulinic acids, dehydrated humulinic acids, beta acids, reduced derivatives of beta acids, oxidized derivatives of beta acids, or combinations thereof.

[0131] In some aspects, the compositions disclosed herein comprise one or more beta acid derivatives, one or more salts of beta acids, or combinations thereof.

[0132] In some aspects, the beta acid derivatives are selected from lupulones,

hexahydrolupulones, deoxyhumulones, deoxytetrahydrohumulones, hulupones, dihydro- hulupones, tetrahydrohulupones, hexahydrohulupones, hulupinic acids, lupoxes, lupdoxes, lupdeps, lupdols, tricyclo-oxylupulones, and combinations thereof.

[0133] In some aspects, the compositions disclosed herein comprise one or more alpha acid derivatives.

[0134] In some aspects, the alpha acid derivatives are selected from humulones (alpha acids), dihydrohumulones, tetrahydrohumulones, humulohydroquinones,

humuloquinones, humulinones, tricyclodehydroisohumulones, abeoisohumulones, isohumulones, dihydroisohumulones, tetrahydroisohumulones, hexahydroisohumulones, alloisohumulones, humulinic acids, isomers of humulinic acids, dihydrohumulinic acid and derivatives, oxyhumulinic acids, dehydrohumulinic acids and derivatives, dehydrated humulinic acids, and combinations thereof.

[0135] In one aspect, the compositions disclosed herein comprise at least about 0.1 g/L, at least about 0.2 g/L, at least about 0.3 g/L, at least about 0.4 g/L, at least about 0.5 g/L, at least about 0.6 g/L, at least about 0.7 g/L, at least about 0.8 g/L, at least about 0.9 g/L, at least about 1 g/L, at least about 2 g/L, at least about 3 g/L, at least about 4 g/L, at least about 5 g/L, at least about 6 g/L, at least about 7 g/L, at least about 8 g/L, at least about 9 g/L, at least about 10 g/L, at least about 11 g/L, at least about 12 g/L, at least about 13 g/L, at least about 14 g/L, at least about 15 g/L, at least about 16 g/L, at least about 17 g/L, at least about 18 g/L, at least about 19 g/L, or at least about 20 g/L alpha acids. [0136] In one aspect, the miticide compositions disclosed herein composition comprise at least about 0.1 g/L, at least about 0.2 g/L, at least about 0.3 g/L, at least about 0.4 g/L, at least about 0.5 g/L, at least about 0.6 g/L, at least about 0.7 g/L, at least about 0.8 g/L, at least about 0.9 g/L, at least about 1 g/L, at least about 2 g/L, at least about 3 g/L, at least about 4 g/L, at least about 5 g/L, at least about 6 g/L, at least about 7 g/L, at least about 8 g/L, at least about 9 g/L, at least about 10 g/L, at least about 11 g/L, at least about 12 g/L, at least about 13 g/L, at least about 14 g/L, at least about 15 g/L, at least about 16 g/L, at least about 17 g/L, at least about 18 g/L, at least about 19 g/L, or at least about 20 g/L iso alpha acids.

[0137] In a particular aspect, the compositions disclosed herein comprise a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) comprising a mixture of alpha acid derivatives (e.g., iso alpha acids) and beta acid derivatives.

5. Formulations comprising Bulnesia sp. plant extracts and Humulus sp. plant extracts

[0138] The present disclosure also provides formulations comprising at least one

Bulnesia sp. plant extract (e.g., an extract, a purified fraction, a partially purified fraction, or a combination thereof) and at least one Humulus sp. plant extract (e.g., an extract, a purified fraction, a partially purified fraction, or a combination thereof). In some aspect, these formulations are used as miticides. In some aspects, the Bulnesia sp. and the Humulus sp. plant extracts in the formulation function synergistically as miticides. In other aspects, the formulations disclosed herein can be used, for example, as insecticides, nematicides, bactericides, or fungicides.

[0139] In some aspects, the Bulnesia sp. plant extract in the formulation is obtained, e.g., from Bulnesia sarmientoi. In some aspects, the Humulus sp. plant extract in the formulation is obtained, e.g., from Humulus lupulus. In some aspects, the Humulus sp. plant extract comprises iso alpha acids, which can be, e.g., from plant origin, synthetic, semisynthetic, or a combination thereof. In some aspects, the concentration of iso alpha acids in the Humulus sp. plant extract is at least 75%.

[0140] In a specific aspect, the formulations disclosed herein comprise a miticide

composition comprising: (a) a Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) wherein the Bulnesia sp. plant extract is obtained, e.g., from Bulnesia sarmientoi; and

(b) a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) wherein the Humulus sp. plant extract is obtained, e.g., from Humulus lupulus, and wherein the iso alpha acid content in the Humulus sp. plant extract is at least about 75% (i.e., it is at least about 75% pure),

[0141] In some aspects, the compositions disclosed herein comprise a Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) and a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof), and further comprise excipients.

[0142] The compositions disclosed herein can comprise any acceptable excipients known in the art for their intended use. For example, a composition comprising (i) & Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) wherein the Bulnesia sp. plant extract is obtained, e.g., from Bulnesia sarmientoi; and (ii) a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) wherein the Humulus sp. plant extract is obtained, e.g., from Humulus lupulus, for use as an insecticide, would comprise insecticidally acceptable excipients known the art. Similarly, a composition for use as a nematicide, bactericide, or fungicide would comprise acceptable excipients known in the art for their intended purpose.

[0143] The compositions disclosed herein can be formulated including any excipients and in any of the formats know in the art {e.g., powders, solutions, etc.).

[0144] In some aspects, the excipients include, for example, acacia gum, acacia gum

modified with ocetenyl succinic anhydride, acetylated monoclycerides, acetylated tartaric acid esters of mono- and di- glycerides, agar, algin, alginic acid, ammonium alginate, ammonium carrageenan, ammonium furcelleran, ammonium salt of phosphorylated glyceride, arabino-galactan, baker's yeast glycan, calcium alginate, calcium carbonate, calcium carrageenan, calcium citrate, calcium furcelleran, calcium gluconate, calcium glycerophosphate, calcium hypophosphite, calcium lignosulphonate, calcium phosphate, calcium sulphate, calcium tartrate, carboxymethyl cellulose, carob bean gum,

carrageenan, cellulose gum, citric acid esters of mono- and di-glycerides, gurcelleran, gelatin, gellan gum, guar gum, gum Arabic, hydroxylated lecithin, hydroxypropyl cellulose, irish moss gelose, karaya gum, lactylated mono- and di-glycerides, lactylic esters of fatty acids, lecithin, locust bean gum, magnesium chloride, methylcellulose, methyl ethyl cellulose, monoglycerides, mono- and di-glycerides, monosodium salts of phosphorylated mono- and di-glycerides, oat gum, pectin, polyglycerol esters of interesterified castor oil fatty acids, polyoxyethylene, sorbitan monooleate,

polyoxyethylene, sorbitan monostearate, polyoxyethylene, sorbitan tristearate, polyoxyethylene stearate, potassium alginate, potassium carrageenan, potassium chloride, potassium citrate, potassium furcelleran, potassium phosphate (dibasic), propylene glycerol alginate, propylene glycol ether of methylcellulose, propylene glycol mono fatty acid esters, sodium acid pyrophosphate, sodium alginate, sodium aluminum phosphate, sodium carboxymethyl cellulose, sodium carrageenan, sodium cellulose glycolate, sodium chloride, sodium citrate, sodium furcelleran, sodium gluconate, sodium hexametaphosphate, sodium phosphate (dibasic), sodium phosphate (monobasic), sodium phosphate (tribasic), sodium potassium tartrate, sodium pyrophosphate (tetrabasic), sodium stearoyl-2-lactylate, sodium stearate, sodium tartrate, sodium tripolyphosphate, sorbitan monostearate, sorbitan trioleate, sorbitan tristearate, stearyl monoglyceridyl citrate, sucrose esters of fatty acids, tragacanth gum, xanthan gum, calcium silicate, belite calcium orthosilicate, grammite, micro-cell, silene, silicic acid calcium salt, and combinations thereof. In some aspects, the excipient is calcium lignosulphonate.

[0145] In some aspects, the excipient is carboxymethyl cellulose. In some aspects, the excipient is sodium chloride. In some aspects, the excipient is calcium silicate. In some aspects, the use of excipients in a miticide composition is for the purpose other than the miticide effect, for example, a stabilizing compound, a buffering compound, an emulsifying compound, a bulking agent or a combination thereof.

[0146] In some aspects, the compositions disclosed herein can be formulated in a solid form. In some aspects, the solid form comprises, for example, powder, pellet, or granule formulations. In some aspects, the powder, pellet, or granule formulations are dispersible. In some aspects, the dispersible powder, pellet, or granule formulations are water- dispersible. In some aspects, the solid form formulation is, for example, a pellet, a granule, a powder, a dry flowable, a bait, dust, a nanoencapsulated formulation, a microencapsulated formulation, or a combination thereof. In some aspects, the powder formulations are wettable powder formulations.

[0147] In some aspects, the compositions disclosed herein can be formulated in liquid form. In some aspects, the liquid form is, for example, a liquid concentrate formulation, an emulsifiable concentrate, an emulsion, a suspension, a liquid flowable, a gel, a ready- to-use formulation, an oil dispersion, an aerosol formulation, or combinations thereof. In some aspects, the liquid concentrate is an ultra-low- volume concentrate formulation. In some aspects, the liquid formulations disclosed herein are emulsifiable concentrates (EC), emulsions in water (EW), or oil dispersions (OD).

[0148] In some aspects, the emulsifiable concentrate (EC) contains an organic solvent and a surfactant in addition to the active ingredients. In some aspects, the liquid formulation is an emulsion in water (EW).

[0149] In some aspects, the organic solvent can be replaced by an oil (e.g., vegetable or mineral), lowering the volatilization of volatile compounds -VOC- and reducing the risks of phytotoxicity and flammability. These formulations are known as Oil dispersions (OD). In some aspects, the OD formulation uses an oil, for example, a vegetable oil as solvent. In some aspects, the OD formulation comprises additional components, e.g., co- solvents with high polarity and/or adjuvants. In some aspects, such co-solvents and/or adjuvants confer sticky (adhering) properties to the sprayed plants.

[0150] In some aspects, the EC, EW and OD formulations disclosed herein can maintain the active ingredients in solution in a temperature range from about -20°C to 70°C. In other aspects, the EC, EW and OD formulations disclosed herein can maintain the active ingredients in solution in a temperature range from about -10°C to 54°C. In some aspects, EC, EW and OD formulations disclosed herein can maintain the active ingredients without aggregation (e.g., crystallization) in a temperature range from about -20°C to 70°C. In other aspects, EC, EW and OD formulations disclosed herein can maintain the active ingredients without aggregation (e.g., crystallization) in a temperature range from about -10°C to 54°C.

[0151] In some aspects, EC, EW and OD formulations disclosed herein can maintain the active ingredients without aggregation (e.g., crystallization) at about -20°C, at about - 19°C, about -18°C, at about -17°C, about -16°C, at about -15°C, about -14°C, at about - 13°C, about -12°C, at about -11°C, about -10°C, at about -9°C, about -8°C, about -7°C, about -6°C, about -5°C, about -4°C, about -3°C, about -2°C, about -1°C, about 0°C, about 1°C, about 2°C, about 3°C, about 4°C,about 5°C, about 10°C, about 15°C, about 20°C, about 25°C, about 30°C, about 35°C, about 40°C, about 45°C, about 50°C, about 55°C, about 60°C, about 65°C or about 70°C.

[0152] In some aspects, EC, EW and OD formulations disclosed herein can maintain the active ingredients in solution at about -20°C, at about - 19°C, about -18°C, at about - 17°C, about -16°C, at about -15°C, about -14°C, at about -13°C, about -12°C, at about - 11°C, about -10°C, at about -9°C, about -8°C, about -7°C, about -6°C, about -5°C, about - 4°C, about -3°C, about -2°C, about -1°C, about 0°C, about 1°C, about 2°C, about 3°C, about 4°C,about 5°C, about 10°C, about 15°C, about 20°C, about 25°C, about 30°C, about 35°C, about 40°C, about 45°C, about 50°C, about 55°C, about 60°C, about 65°C or about 70°C.

[0153] In some aspects, the EC, EW and OD formulations disclosed herein comprise as active ingredients a combination of (i) a Humulus sp. plant extract, e.g., a Humulus lupulus plant extract, and (ii) a Bulnesia sp. plant extract, e.g., a Bulnesia sarmientoi plant extract, wherein the combination acts synergistically as a miticide, wherein the Humulus sp. plant extract concentration in the formulation ranges from 0 g/L to 1.5 g/L, and whereas the concentration of Bulnesia sp. plant extract concentration in the formulation ranges from 0 g/L to 5 g/L.

[0154] In some aspects, the Humulus sp. plant extract concentration in the formulation is at least about 0.1 g/L, at least about 0.2 g/L, at least about 0.3 g/L, at least about 0.4 g/L, at least about 0.5 g/L, at least about 0.6 g/L, at least about 0.7 g/L, at least about 0.8 g/L, at least about 0.9 g/L, at least about 1 g/L, at least about 1.1 g/L, at least about 1.2 g/L, at least about 1.3 g/L, at least about 1.4 g/L, at least about 1.5 g/L, at least about 1.6 g/L, at least about 1.7 g/L, at least about 1.8 g/L, at least about 1.9 g/L, at least about 2 g/L, at least about 2.1 g/L, at least about 2.2 g/L, at least about 2.3 g/L, at least about 2.4 g/L, or at least about 2.5 g/L. In some aspects, the Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) is an extract prepared according to the method of Example 2.

[0155] In some aspects, the Bulnesia sp. plant extract concentration in the formulation is at least about 0.1 g/L, at least about 0.2 g/L, at least about 0.3 g/L, at least about 0.4 g/L, at least about 0.5 g/L, at least about 0.6 g/L, at least about 0.7 g/L, at least about 0.8 g/L, at least about 0.9 g/L, at least about 1 g/L, at least about 1.1 g/L, at least about 1.2 g/L, at about 1.3 g/L, at least about 1.4 g/L, at least about 1.5 g/L, at least about 1.6 g/L, at about 1.7 g/L, at least about 1.8 g/L, at least about 1.9 g/L, at least about 2 g/L, at about 2.1 g/L, at least about 2.2 g/L, at least about 2.3 g/L, at least about 2.4 g/L, at about 2.5 g/L, at least about 2.6 g/L, at least about 2.7 g/L, at least about 2.8 g/L, at about 2.9 g/L, at least about 3 g/L, at least about 3. 1 g/L, at least about 3.2 g/L, at about 3.3 g/L, at least about 3.4 g/L, at least about 3.5 g/L, at least about 3.6 g/L, at about 3.7 g/L, at least about 3.8 g/L, at least about 3.9 g/L, at least about 4 g/L, at about 4.1 g/L, at least about 4.2 g/L, at least about 4.3 g/L, at least about 4.4 g/L, at about 4.5 g/L, at least about 4.6 g/L, at least about 4.7 g/L, at least about 4.8 g/L, at about 4.9 g/L, at least about 5 g/L, at least about 5. 1 g/L, at least about 5.2 g/L, at about 5.3 g/L, at least about 5.4 g/L, at least about 5.5 g/L, at least about 5.6 g/L, at about 5.7 g/L, at least about 5.8 g/L, at least about 5.9 g/L, at least about 6 g/L, at about 6.1 g/L, at least about 6.2 g/L, at least about 6.3 g/L, at least about 6.4 g/L, at about 6.5 g/L, at least about 6.6 g/L, at least about 6.7 g/L, at least about 6.8 g/L, at about 6.9 g/L, at least about 7 g/L, at least about 7. 1 g/L, at least about 7.2 g/L, at about 7.3 g/L, at least about 7.4 g/L, or at least about 7.5 g/L. In some aspects, the

Bulnesia sp. plant extract (e.g., a. Bulnesia sarmientoi plant extract) is an extract prepared according to the method of Example 3.

[0156] In some aspects, a formulation comprising a composition disclosed herein

comprises a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) wherein the Humulus sp. plant extract at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%), at least about 14%, or at least about 15% of the formulation (w/w or w/v). In some aspects, the Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) is an extract prepared according to the method of Example 2.

[0157] In some aspects, a formulation comprising a composition disclosed herein

comprises a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) wherein the Bulnesia sp. plant extract at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%), at least about 18%, at least about 19%, at least about 20%, at least about 21%, at least about 22%, at least about 23%, at least about 24%, at least about 25%, at least about 26%o, at least about 27%, at least about 28%, at least about 29%, at least about 30%, at least about 31%, at least about 32%, at least about 33%, at least about 34%, at least about 35%o, at least about 36%, at least about 37%, at least about 38%, at least about 39%, at least about 40%, at least about 41%, at least about 42%, at least about 43%, at least about 44%o, at least about 45%, at least about 46%, at least about 47%, at least about 48%, at least about 49%, or at least about 50% of the formulation (w/w or w/v). In some aspects, the Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) is an extract prepared according to the method of Example 3.

[0158] In some aspects, the EC, EW and OD formulations disclosed herein comprise a

Humulus sp. plant extract obtained, for example, from Humulus lupulus wherein the concentration of iso alpha acids is between 20% and 99% (w/v). In other aspects, the concentration of iso alpha acids is between 20% and 80% (w/v). In some aspects, the concentration of iso alpha acids is at least about 20%, at least about 25%, at least about 30%o, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%o, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% (w/v). In a particular aspect, the concentration of iso alpha acids is between 70% and 80% (w/v). In another particular aspect, the concentration of iso alpha acids is about 75% (w/v).

[0159] The trade names presented in TABLES 1-6 are not limiting and are provided only as examples corresponding to the chemical families presented in the right column of each table. In other words, the disclosure of ALKAMULS™ T/85-V as an ethoxylated sorbitan trioleate in TABLE 1 does not mean that ALKAMULS™ T/85-V is the only ethoxylated sorbitan trioleate contemplated for the EW formulations disclosed herein. Instead, it indicates that any ethoxylated sorbitan trioleate can be used in the EW compositions disclosed herein, and that in specific formulations such ethoxylated sorbitan trioleate could be ALKAMULS™ T/85-V. The same interpretation would apply to every single entry in TABLES 1-6.

[0160] Suitable surfactants, rheology modifiers, and solvents for use in the EW

formulations disclosed herein are shown in TABLES 1, 2, and 3, respectively. TABLE 1. Surfactants for use in EW formulations

TABLE 2. Rheology modifiers for use in EW formulations

TABLE 3. Solvents for use in EW formulations

Trade name Chemical family

Acetophenone Phenyl Methyl Ketone

AMESOLV ADM A 10™ N,N~Di emethy 1 decanamide

AMESOLV ADMA 810™ N,N~Dimethyi octanarnide and N,N~Diemethyi decanamide AMOIL BH™ Rapeseed Oil refined

AM OIL SUN F™ Sunflower Oil refined

Com Oil Cora Oil refined Di m ethyl sulph oxi de Di methy 1 s u lph oxi de

PHYTOROB 810.01™ Methyl Ester C8-C 10

PHYTOROB 926.65™ Methyl Oleate

Propylene Carbonate Propylene Carbonate

RHODIASOLV GREEN 21™ Solvents mixture

RHODIASOLV GREEN 25™ Solvents mixture

Soja Oil Soybean Oil refined

Glycerine Glycerine

[0161] Suitable surfactants, rheology modifiers, and solvents for use in the OD formulations disclosed herein are shown in TABLES 4, 5, and 6, respectively.

TABLE 4. Surfactants for use in OD formulations

Trade name Chemical family

ALKAMULS VO/2003™ Ethoxylated Fatty Acids

RHODASURF LA/30™ Ethoxylated Alcohol

RHODASURF LA/40™ Ethoxylated Alcohol

TWEEN 20™ Polysorbate 20

TWEEN 80™ Polysorbate 80

TWEEN 85™ Polysorbate 85

SPAN 20™ Sorbitan laurate

SPAN 80™ Sorbitane monooleate

SPAN 85™ Sorbitan stearate

TABLE 5. Rheology modifiers for use in OD formulations

Trade name Chemical family

ATTAGEL® 40 Attapulgite gelling grade

ΒΕΝΤΟΝΠΈ® AG/8 Montmorillonite and other Al-silicates

TABLE 6. Solvents for use in OD formulations

Trade name Chemical family

Oil BH® Rapeseed Oil refined

AMOIL SUN® F Sunflower Oil refined

Corn Oil Corn Oil refined

PHYTOROB© 810.01 Methy ^j Ester C8-C 10

PHYTOROB® 926.65 Methyl Oleate

Soja Oil Soybean Oil refined

Canola oil Canola oil

Glycerine Glycerine

Castor Oil

Mineral Oil

Sesame Oil [0162] In some aspects, the compositions disclosed can further comprise a carrier. In some aspects, the carrier is, for example, a neutral carrier edible, for example, for mites. In some aspects, the carrier further comprises, for example, a dye, a pigment, a safety additive agent, an attractant, bait, an agent improving rain-resistance, or combinations thereof.

[0163] The term "bait" as used herein refers to a products readily ingested by a pest (e.g., a mite, insect, nematode, etc.). In some aspects, the bait is, for example, agar, potato dextrose agar, sugar beet, gelatin, oil cake, pet food, wheat, soya, oats, corn, rice, fruits, fish by-products, sugars, coated vegetable and cereal seeds, casein, blood meal, bone meal, yeast, paper products, clays, fats, a variety of cereals, including wheat cereal or combinations thereof.

[0164] Baits, in general, are agrochemical formulation, in which the active ingredient is mixed with some inert substances with attractant effect, such as food, syrups, and pheromones among others. This kind of formulations can be solids, liquids, gels or arranged physical traps.

[0165] In some aspects, the formulations disclosed herein comprise a Bulnesia sp. plant extract {e.g., a crude extract, or an extract enriched in some compounds) and a Humulus sp. plant extract (e.g., a crude extract, or an extract enriched in some compounds, for example an extract enriched in iso alpha acids), and additional products such as inert components, attractants, water repellents, binders, preservatives, surfactants, rheology modifiers, emulsifiers, adjuvants, stabilizers, or combinations thereof.

[0166] In some aspects, a product added to the active ingredients (Bulnesia sp. plant extract, and Humulus sp. plant extract) in a formulation disclosed herein can have multiple functions, e.g., glycerin can function as both stabilizer and adjuvant, or

TWEEN™ can function as both emulsifier and surfactant.

6. Use compositions comprising Bulnesia sp. and Humulus sp. plant extracts as miticides

[0167] The present disclosure also provides methods to control, prevent, or treat mite infestations by using any of the compositions disclosed herein, for example, a

composition comprising a Bulnesia sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof) and a Humulus sp. plant extract (e.g., an extract, or a purified fraction or partially purified fraction thereof), wherein the Bulnesia sp. plant extract (obtained, e.g., from Bulnesia sarmientoi) and the Humulus sp. plant extract (obtained, e.g., from Humulus lupulus) function synergistically as miticides

[0168] The compositions and formulations disclosed herein can also be used to as

repellants. The miticide compositions disclosed herein can act by contact, by ingestion, or simultaneously by both mechanisms of action. The disclosed compositions can have toxic effects, i.e., they can interfere with the mite metabolism once they make contact with the mite's tissue or when treated plant tissue is ingested by the mite. The disclosed

compositions can also act as anti-feedants, i.e., the disclosed compositions can prevent damage cause by feeding on plant tissue (e.g., leaves, fruits, seeds, etc.) not through toxic mechanism but by discouraging mites from feeding on plant tissue by making it non- palatable. Thus, in some aspects, the disclosed compositions prevent damage caused by mites through a toxicity-based mechanism of action. In other aspects, the disclosed compositions prevent damage caused by mites through an anti-feeding-based mechanism of action. In other aspects, the disclosed compositions prevent damage caused by mites by both mechanisms of action.

[0169] In a specific aspect, the miticide composition used in the methods disclosed herein comprises:

(a) a Humulus sp. plant extract from Humulus lupulus, wherein said extract is enriched in iso alpha acids; and

(b) a Bulnesia sp. plant extract from Bulnesia sarmientoi,

wherein the Bulnesia sp. plant extract and the Humulus sp. plant extract act synergistically as miticides. In some specific aspects, the Humulus sp. plant extract is prepared according to the method of Example 2 and/or the Bulnesia sp. plant extract is prepared according to the method of Example 3.

[0170] Mite infestations can occur in agricultural environments, horticultural

environments, gardens, or any combination thereof. Accordingly, the miticide

compositions disclosed herein can be applied, for example, in agricultural environments, horticultural environments, gardens, or any combination thereof. In some aspects, the miticide compositions can be applied to animals, including humans.

[0171] The term "treat" as used herein to refer to miticide treatments encompasses any beneficial effect derived from the application of an effective amount of any of the compositions disclosed herein, including the eradication, control, or prevention of mite infestations. Eradication refers to the complete removal of mites from a certain space or plant population. Similarly, when referring to nematicide treatments, insecticide treatments, bactericide treatments, fungicide treatment, etc., the term treat refers to any beneficial effect derived from the application of an effective amount of any of the compositions disclosed herein, including the eradication, control, or prevention of infestation or infection by the target organism.

[0172] As used herein, the term "control" and grammatical variants of the term refers to actions that result in a reduction in the number of mites or damage caused by mites in a certain area after the application of a miticidally effective amount of any of the miticide compositions disclosed herein. Such actions can be for example an increase in mite mortality, a reduction in mite longevity, a reduction in mite reproduction rate,

physiological and/or metabolic changes that reduce the ingestion rate of plant material by the mites, repellant effect that discourages the movement of mites to a certain area or their movement onto treated surfaces or their attachment to treated surfaces, etc. Similarly, when referring to nematicide treatments, insecticide treatments, bactericide treatments, fungicide treatment, etc., the term control refers to actions that result in a reduction in the number of target organism or damage caused by the target organism in a certain area after the application of an effective amount of any of the compositions disclosed herein..

[0173] In some aspects, the compositions and/or formulations disclosed herein can be applied to a certain area, in the absence of a mite infestation (or any other pest infestation, e.g., a nematode or insect infestation), to prevent the occurrence of mite infestations (or any other pest infestation, e.g., a nematode or insect infestation), or to prevent mite re- infestations (or any other pest re-infestation, e.g., a nematode or insect infestation) after an area has been cleared of mites (or any other pests, e.g., nematodes or insects) after the application of any of the compositions disclosed herein or the application of any other pesticide compositions known in the art.

[0174] The phrase "effective amount" as used herein refers to the amount of the a

composition disclosed that, when ingested or sensed by a pest (e.g., when contacting mites), is sufficient to achieve a desired pesticide effect, e.g., killing, repelling, preventing, or any combination thereof.

[0175] For example, concentrations of the compositions disclosed herein, for example, miticide compositions and formulations are, for example, effective for treating the infestation of mites in an afflicted area. The effective amount is, for example, that amount of the compositions disclosed herein, necessary and/or sufficient to perform its intended function within a mite, e.g., to cause expiration, to cause the mite to repel, to prevent the mite from entering, or any combination thereof. An effective amount of the composition can vary according to factors, for example, the amount of the causative agent already present in the mite, the size of the mite, the density of the mite, the developmental stage of the mite, the health of the mite, the species of mite, the host plant of the mite, the location of the mite on the plant, environmental conditions (e.g., humidity, precipitation, wind, temperature, etc.), or any combination thereof.

[0176] One of ordinary skill in the art would be able to study the aforementioned factors and make a determination regarding the effective amount of the compositions disclosed herein without undue experimentation. In some aspects, an in vitro or in vivo assay also can be used to determine an "effective amount" of the compositions disclosed herein.

[0177] In some aspects, the effective amount of the composition of the disclosure (e.g., a miticide composition, insecticide composition, nematicide composition, etc.) preferably eliminates at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%>, at least about 35%, at least about 40%, at least about 45%, at least about 50%), at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%), at least about 97%, at least about 99%, or 100% of the pest (e.g., mites) relative to untreated areas.

[0178] In some aspects, control, prevent, and/or treat pest (e.g., mite) infestation is

reducing the number of pests (e.g., mites). In some aspects, control, prevent, and/or treat pest (e.g., mite) infestation is reducing the amount of pest (e.g., mite) damage.

[0179] In some aspects, control, prevent, and/or treat pest (e.g., mite) infestation is

reducing the number of pests (e.g., mites) in a certain area. In some aspects, control, prevent, and/or treat pest (e.g., mite) infestation is increasing the percent pest (e.g., mite) mortality.

[0180] In some aspects, the number of pests (e.g., mites) is reduced by about 5%, by

about 10%, by about 20%, by about 30%, by about 40%, by about 50%, by about 60%, by about 70%, by about 80%, by about 90% by about 95%, or by about 100%. [0181] In some aspects, the reduction of pest (e.g., mite) damage is by about 5%, by about 10%, by about 20%, by about 30%, by about 40%, by about 50%, by about 60%, by about 70%, by about 80%, by about 90% by about 95%, or by about 100%.

[0182] In some aspects, the number of pest organisms (e.g., mites) reduced in a certain area is about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% about 95%, or about 100%.

[0183] In some aspects, the percent mortality of pest organisms (e.g., mites) is increased by about 5%, by about 10%, by about 20%, by about 30%, by about 40%, by about 50%, by about 60%, by about 70%, by about 80%, by about 90% by about 95%, or by about 100%.

[0184] In some aspects, the method for controlling, treating, and/or preventing pest (e.g., mite) damage to plants according to the compositions disclosed herein is, for example, treating the plants or the locus surrounding the plants with an effective amount (e.g., a miticidally effective amount) of any of the compositions disclosed herein. In some aspects, the locus surrounding the plants can be treated. Such locus can be, for example, an area of known pest (e.g., mite) mite infestation, an area of unknown pest (e.g., mite) infestation, an area with the potential for pest (e.g., mite) infestation or combinations thereof.

[0185] In some aspects, the compositions disclosed herein (e.g., miticide, nematicide or insecticide compositions) can be used to treat a surface, for example, soil, grass, flower beds, sidewalks, driveways, crates, containers, fields, crops, or combinations thereof.

[0186] In some aspects, the soil adjacent to the base of a plant can be treated. In other aspects, the plant itself, for example the stems, leaves, fruits, roots, bark, flowers, buds, etc. can be treated.

[0187] In some aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) can be administered, for example, by spreading, shaking, dispersing, scattering, distributing, strewing, scattering, throwing, casting, littering, low pressure spraying, high pressure spraying, brushing, misting, vaporizing, volatilizing, fogging, fumigating, immersing, injecting, vapor treating, pressure treating, drenching, drip irrigating, atomizing, broadcasting, foaming, or combinations thereof.

[0188] In some aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) can be administered, for example, using a fogger, a sprayer, a mister, a diffusor, a box, an envelope, a paper, gloves, a shovel, a rake, a bait trap, a hose- end sprayer, a hand-powered applicator, a rotary and drop spreader, hand distribution, sprinkler, a container, or combinations thereof.

[0189] In some aspects, the compositions (e.g., miticide, nematicide, or insecticide

compositions) used in the methods disclosed herein are applied according to a certain dose regimen. The phrase "dose regimen" as used herein refers the frequency of administration of the composition (e.g., miticide, nematicide, or insecticide compositions) that is sufficient to achieve pest (e.g., mite, nematode, or insect) control, treatment, prevention, or any combination thereof. In some aspects, the treatment comprises a single dose. In other aspects, the treatment comprises more than one dose.

[0190] In some aspects, the dose regimen consists in daily, weekly, or monthly

applications of the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions). In some cases, one or more doses can be applied in a single day. In some aspects, the dose regimen consists of one daily dose, two daily doses, three daily doses, four daily doses, or more than four daily doses. In some aspect, the dose regimen consists of one weekly dose, two weekly doses, three weekly doses, four weekly doses, five weekly doses, six weekly doses, or more than six weekly doses. In some aspects, the dose regimen consists of one monthly dose, two monthly doses, three monthly doses, four monthly doses, five monthly doses, six monthly doses, seven monthly doses, eight monthly doses, nine monthly doses, ten monthly doses, eleven monthly doses, twelve monthly doses, or more than twelve monthly doses. In some aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) are administered yearly.

[0191] In specific aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) are administered in weekly doses. In other specific aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) are administered in monthly doses. In some aspects, when multiple doses are

administered, the same amount of composition is administered in each dose. In other aspects, the amount of composition administered in each dose varies. For example, in some cases it can be desirable to administer a larger first dose or initial series of doses, followed by maintenance doses which are smaller than the initial (loading) dose or series of doses. In some aspects, when multiple doses are administered, such doses are evenly spaced. For example, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) can be administered once every week. In other cases, multiple doses are not evenly spaced. For example, it can be desirable to administered the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) daily during the first week of treatment, and after the first week of treatment administer the composition only once per week.

The miticide compositions, formulations, and methods disclosed herein can be used to control mites such as Bryobia neopraetiosa (brown lucerne mite), Bryobia praetiosa (brown clover mite), Bryobia rubrioculus (brown fruit tree mite or bryobia mite), Eotetranychus falcatus (cotton green mite), Eotetranychus lewisi (poinsettia spider mite), Eutetranychus orientalis (Lowveld citrus mite or Oriental mite), Meyernychus emeticae (Angolan citrus mite), Mononychellus tanajoa (cassava green mite),

Mononychellus progresivus (cassava green mite), Oligonychus coffeae (red tea mite), Oligonychus mangiferus (mango red mite), Panonychus citri (citrus red mite),

Panonychus ulmi (European red mite), Petrobia latens (brown wheat mite or onion mite), Schizotetranychus asparagi (asparagus mite), Tetranychina harti (oxalis mite), Tetranychus amicus (African spider mite or banana mite), Tetranychus evansi (tobacco spider mite), Tetranychus hydrangeae (hydrangea spider mite), Tetranychus lombardinii (crimson spider mite), Tetranychus ludeni (dark-red spider mite or red-legged spider mite), Tetranychus neocaledonicus (vegetable spider mite), Tetranychus rooyenae (cotton spider mite), Tetranychus turkestani (Turkish spider mite), Tetranychus urticae (common spider mite or two-spotted spider mite), Abacarus hystrix (cereal rust mite), Aceria aloinis (aloe wart mite or aloe gall mite), Aceria barbertoni (Gerbera erineum mite), Aceria cynodoniensis (Bermuda grass stunt mite or grass rosette mite), Aceria diastolus (Cape leadwort leafroll mite), Aceria ficus (fig bud mite), Aceria granati (pomegranate leafroll mite or pomegranate leafcurl mite), Aceria mangiferae (mango bud mite), Aceria merwei (sugarcane mite), Aceria oleae (olive bud mite), Aceria proteae (Protea witches' broom mite), Aceria sheldoni (citrus bud mite), Aceria tulipae (tulip bulb mite or bulb eriophyid mite), Aceria tumisetus (Gazania gall mite), Aculops lycopersici (tomato rust mite), Aculus fockeui (peach silver mite), Calacarus capsica (pepper purple mite), Calacarus carinatus (purple tea mite), Calacarus citrifolii (grey citrus mite or citrus grey mite), Colomerus spathodeae (African flame erineum mite or tulip tree erineum mite), Colomerus vitis (grape gall mite, grape bud mite, grape erinose mite or grape blister mite), Epitrimerus pyri (pear rust mite), Eriophyes pyri (pear bud mite or pear leaf blister mite), Phyllocoptruta oleivora (citrus rust mite), or Phytoptus hedericola (ivy bud mite).

[0193] In some aspects, the compositions disclosed herein can be used to treat, prevent, or ameliorate leaf miner infestation in plants. Leaf miners are the larva of insects that live in and eats the leaf tissue of plants. The vast majority of leaf-mining insects are moths (Lepidoptera), sawflies (Symphyta, a type of wasp) and flies (Diptera), though some beetles also exhibit this behavior. Leaf miners are regarded as pests by many farmers and gardeners as they can cause damage to agricultural crops and garden plants, and can be difficult to control with insecticide sprays as they are protected inside the plant's leaves. In some aspects, the leaf miner belongs to the genus Liriomyza, e.g., Liriomyza huidobrensis, Liriomyza trifolii (American serpentine leaf miner), or Liriomyza sativae (vegetable leaf miner). In some aspects, the leaf miner belongs to one of the genera in the Agromyzidae family, e.g., Agromyza, Amauromyza, Aulagromyza, Calycomyza, Cecidomyiaceltis, Cerodontha, Chromatomyia, Galiomyza, Geratomyza,

Gymnophytomyza, Haplopeodes, Hexomyza, Indonapomyza, Japanagromyza,

Kleinschmidtimyia, Liriomyza, Melanagromyza, Metopomyza, Napomyza, Nemorimyza, Ophiomyia, Penetagromyza, Phytobia, Phytoliriomyza, Phytomyza, Pseudoliriomyza, Pseudonapomyza, Ptochomyza, Selachops, Tropicomyia, or Xeniomyza. In some aspects, the leaf miner belongs to the genus Pegomya, e.g., Pegomya hyoscyami (beet leaf miner or spinach leaf miner). In some aspects, the leaf miner belongs to the genus Phyllocnistis, e.g., Phyllocnistis citrella (citrus leaf miner). In some aspects, the leaf miner is a moth larva from the Douglasiidae family (e.g., belonging to the genera Klimeschia, Protonyctia or Tinagma). In some aspects, the leaf miner is a larva (casterpillar) belonging to the moth Gracillariidae family, e.g., from the genera Acrocercops, Caloptilia, Cameraria, Epicephala, or Phyllonorycter. In a particular aspect, the leaf miner is a horse-chestnut leaf miner, Cameraria ohridella.

[0194] In some aspects, the compositions disclosed herein can be used to treat, prevent, or ameliorate white fly infestation in plants and/or prevent infections caused by white flies. Whiteflies are small Hemipterans that typically feed on the undersides of plant leaves. They comprise the family Aleyrodidae, the only family in the superfamily Aleyrodoidea. More than 1550 species have been described. In warm or tropical climates and especially in greenhouses, whiteflies present major problems in crop protection. Worldwide economic losses are estimated at hundreds of millions of dollars annually. Prominent pest species include: Aleurocanthus woglumi, citrus blackfly, which, in spite of its color, is a whitefly that attacks citrus; Aleyrodes proletella, cabbage whitefly, is a pest of various Brassica crops; Bemisia argentifolii, silverleaf whitefly, is a pest of many agricultural and ornamental crops; Trialeurodes vaporariorum, greenhouse whitefly, a major pest of greenhouse fruit, vegetables, and ornamentals. Accordingly, in some aspects of the present disclosure, the white fly belongs to the genera Aleurocanthus, Aleyrodes, Bemisia, or Trialeurodes.

[0195] White flyes can be disease vectors, accordingly, control of white fly populations by administering the compositions disclosed herein can be used to prevent infections caused/spread by white flyes, or to contain or ameliorate such infections. The most prominent disease vectors among the Aleyrodidae are a species complex in the genus Bemisia. Bemisia tabaci and B. argentifolii transmit African cassava mosaic, bean golden mosaic, bean dwarf mosaic, bean calico mosaic, tomato yellow leaf curl, tomato mottle, and other Begomoviruses, in the family Geminiviridae. The worldwide spread of emerging biotypes, such as B. tabaci biotype B, also known as, 'B. argentifolii', and a new biotype Q, continue to cause severe crop losses which are expected to increase, demanding matching increases in pesticide use on many crops (tomatoes, beans, cassava, cotton, cucurbits, potatoes, sweet potatoes). Whiteflies and the viruses they carry can infect many host plants, including agricultural crops, palms, and weeds. Whitefly control is difficult and complex, as whiteflies rapidly develop resistance to chemical pesticides. Accordingly, the compositions and methods disclosed herein can be used to treat white fly infestations by white flies that have become resistant to conventional pesticides.

[0196] The compositions disclosed herein (e.g., miticide, nematicide, or insecticide

compositions), formulations, and methods disclosed herein can be used to treat, prevent, or ameliorate pest infestations in plants, for example ornamental plants or crops (e.g., lettuce, cabbage, strawberries, beans, aromatic herbs, cauliflower, or flowers). Suitable target crops for control of pests using the compositions, formulations and methods disclosed herein include, for example cereals (such as wheat, barley, rye, oats, rice, maize or sorghum); beet (such as sugar or fodder beet); fruit (such as pome fruit, stone fruit, apples, pears, plums, peaches, almonds, cherries, strawberries, raspberries or blackberries); legumes (such as beans, lentils, peas or soya beans); oil crops (such as oil seed rape, mustard, poppies, olives, sunflowers, coconuts, castor, cacao or peanuts);

marrows (such as pumpkins, cucumbers or melons); fiber plants (such as cotton, flax, hemp or jute); citrus fruits (such as oranges, lemons, grapefruits or tangerines); vegetables (such as spinach, lettuce, asparagus, cabbage species, carrots, onions, tomatoes, potatoes, or capsicums); laurels (such as avocado, Cinnamonium or camphor); and tobacco, nuts, coffee, egg plants, sugar cane, tea, pepper, grapevines, hops, the banana family, latex plants and ornamentals.

[0197] For example the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) can be used on any of the following ornamental species:

Ageratum, Alonsoa, Anemone spp., Anisodontea capsenisis, Anthemis, Antirrhinum, Rhododendron spp., Begonia spp. (eg. B. elatior, B. semperflorens, B. n^reux),

Bougainvillea spp., Brachycome spp., Calceolaria, Capsicum annuum, Catharanthus roseus, Ornamental Brassica, Canna spp., Chrysanthemum, Cineraria spp. (C. maritime), Crassula coccinea, Cuphea ignea, Dicentra spectabilis, Dorotheantus, Eustoma grandiflorum, Forsythia, Fuchsia spp., Geranium Gnaphalium, Gomphrena globosa, Heliotropium, Helianthus, Hibiscus, Hortensia, Hosta, Hypoestes phyllostachya,

Impatiens spp. (Walleriana), Iresines, Kalanchoe spp., Lantana camara, Lavatera trimestris, Leonotis leonurus, Lilium, Mesembryanthemum, Mimulus, Nemesia, Tagetes, Dianthus spp. (carnation), Canna, Oxalis, Bellis, Pelargonium spp. (P. peltatum, P.

Zonale), Viola spp. (pansy), Petunia, Plecthranthus, Poinsettia, Parthenocissus spp. (P. Quinquefolia, P. Tricuspidata), Primula, Ranunculus, Rosa spp. (rose), Salvia, Scaevola aemola, Schizanthus wisetonensis, Solanum, SurPnia, Tagetes spp., Nicotinia, Verbena spp., Zinnia spp. and other bedding plants. Preferred within this class of ornamental crops are Viola, Petunia, Begonia, Impatiens, Geranium (including from seeds and cuttings), Chrysanthemum (including from cuttings), Rosa (including pot plants and from cuttings), Poinsettia, Ranunculus, Fuchsia, Salvia and Hortensia.

[0198] The compositions disclosed herein (e.g., miticide, nematicide, or insecticide

compositions), formulations and methods disclosed herein are also suitable for the protection of plant propagation material, for example seed, such as fruits, tubers or kernels, from mites. The propagation material can be treated with the composition prior to planting, for example by soaking, spraying or coating seed prior to sowing. Alternatively, the compositions can be applied directly to the locus at which the propagation material is to be planted (for example onto the ground, into a seed furrow, or into pot plant growing media). The compositions, formulations, and methods disclosed herein can also be used to protect stored products from pest.

[0199] In some aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) and formulations disclosed herein can be applied by foliar spray. An exemplary application by foliar spray comprises diluting the composition in an appropriate amount of a solvent, e.g., water, for example at 5 to 10 grams per liter. The diluted composition would then be applied by foliar spraying according to a

predetermined schedule, for example, on certain days (e.g., every two weeks), one or more times a day (e.g., once a day), and at the same time or different times during the day (e.g., early in the morning or the end of the day). The amount of composition can vary depending on the incidence of the infestation. For example, if the infestation is high, the dosage can be escalated and/or the frequency can be increased (e.g., applying the composition weekly instead of biweekly).

[0200] In some aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) can be used for drench applications. Drench applications are particularly appropriate to control extremely high populations of pest (e.g., mites). In these cases, the compositions disclosed herein can be diluted in a suitable solvent and applied directly to the soil to capacity. In some aspects, drench applications can be complemented with foliar application.

[0201] In some aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) are applied at about 0.1 kg/ha, about 0.2 kg/ha, about 0.3 kg/ha, about 0.4 kg/ha, about 0.5 kg/ha, about 0.6 kg/ha, about 0,7 kg/ha, about 0.8 kg/ha, about 0.9 kg/ha, about 1 kg/ha, about 1.1 kg/ha, about 1.2 kg/ha, about 1.3 kg/ha, about 1.4 kg/ha, about 1.5 kg/ha, about 1.6 kg/ha, about 1.7 kg/ha, about 1.8 kg/ha, about 1.9 kg/ha, about 2.0 kg/ha, about 2.1 kg/ha, about 2.2 kg/ha, about 2.3 kg/ha, about 2.4 kg/ha, about 2.5 kg/ha, about 2.6 kg/ha, about 2.7 kg/ha, about 2.8 kg/ha, about 2.9 kg/ha, about 3.0 kg/ha, about 3.1 kg/ha, about 3.2 kg/ha, about 3.3 kg/ha, about 3.4 kg/ha, about 3.5 kg/ha, about 3.6 kg/ha, about 3.7 kg/ha, about 3.8 kg/ha, about 3.9 kg/ha, about 4.0 kg/ha, about 4.1 kg/ha, about 4.2 kg/ha, about 4.3 kg/ha, about 4.4 kg/ha, about 4.5 kg/ha, about 4.6 kg/ha, about 4.7 kg/ha, about 4.8 kg/ha, about 4.9 kg/ha, about 5.0 kg/ha, about 5.1 kg/ha, about 5.2 kg/ha, about 5.3 kg/ha, about 5.4 kg/ha, about 5.5 kg/ha, about 5.6 kg/ha, about 5.7 kg/ha, about 5.8 kg/ha, about 5.9 kg/ha, or about 6.0 kg/ha. In some particular aspects, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) are applied at more than 6 kg/ha.

[0202] In some aspects, the miticide compositions disclosed herein can be used to treat or prevent bacterial plant infections in which mites are the transmission vector by applying the miticide compositions disclosed herein. For example, Tetranychus urticae transmits Acidovorax citrulli, causal agent of bacterial fruit blotch of watermelon.

[0203] After several case reports of spider mite-induced asthma and allergy, cross- sectional surveys have demonstrated that spider mites of the genera Tetranychus and Panonychus are important allergens in the development of asthma and rhinitis in fruit farmers. Epidemiological surveys have also demonstrated that spider mites are common sensitizing allergens that are related to the prevalence of asthma and rhinitis, even in the non-farming population exposed to spider mites. Kim et al. "Spider-mite allergy and asthma in fruit growers" Curr Opin Allergy Clin Immunol. 2(2): 103-7 (2002). For example, recent investigations indicate that the citrus red mite (Panonychus citri) is the most important allergen affecting citrus-cultivating farmers and children living with asthma, allergic rhinitis, or both. Accordingly, the present disclosure provides methods to reduce or prevent sensitization, reduce of prevent allergic reactions (e.g., asthma or rhinitis), reduce or prevent levels of spider mite allergens, or combinations thereof comprising applying a miticide composition disclosed herein.

7. Additional uses of compositions comprising Bulnesia sp. and Humulus sp. plant extracts

[0204] The compositions disclosed herein comprising a Humulus sp. plant extract and a

Bulnesia sp. plant extract, in addition to being used a miticides, can be adapted for other uses. In the context of those uses, the description of these compositions elsewhere in the specification as "miticide" compositions is non-limiting. Thus, a person of skill in the art would understand that in the context of use as an insecticide, a composition described elsewhere in the application as a "miticide composition" could indeed be used as an "insecticide composition" or a "nematicide composition." (i) Pharmaceutical Compositions and their uses

[0205] The compositions disclosed in the present specification comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) can be formulated as pharmaceutical compositions for the treatment of human subjects in need thereof. In some aspects, the compositions comprise pharmaceutically acceptable vehicles or excipients. These pharmaceutical compositions can be used for example in methods to increase gastrointestinal tolerability, to protect skin, to modulate inflammation, etc. See, e.g., U.S. Pat. Nos. US 4148873, US7722903, U.S. Publ. No. US20060013870, which are herein incorporated by reference their entireties. The pharmaceutical compositions disclosed herein, i.e., any of the compositions disclosed in the present specification comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a

Humulus lupulus plant extract) can also be formulated into a product form selected from the group consisting of toothpastes, creams, ointments, lotions, salves, nasal mists, and oral rinses. In a particular aspect, the composition is formulated into a toothpaste.

Accordingly, the present disclosure also provides a method for treating an oral cavity in a subject comprising applying a therapeutically effective of a toothpaste comparing a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract). In some aspects, the method of treating is prophylactic and prevents the onset of tooth decay. In another aspect, the method of treating is in response to tooth decay, gingivitis, infection, or oral disease. See, e.g., U.S. Publ. No. US20070248549, which is herein incorporated by reference in its entirety.

(ii) Sunscreens

[0206] The present disclosure provides a sunscreen compositions comprising any of the compositions disclosed in the present specification comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a

Humulus lupulus plant extract). In some aspects, the compositions comprise cosmetically acceptable vehicles or excipients. These compositions can be used to absorb ultraviolet radiation, and therefore to protect skin from damage caused by that ultraviolet radiation. Accordingly, these composition can be used as sunscreening compositions to protect said skin from erythema-producing sunlight radiation. These compositions can also be used to promote tanning. See, e.g., U.S. Pat. No. US 4148873, which is herein incorporated by reference in its entirety.

(in) Preservatives and Antibacterials

[0207] The present disclosure provides also antimicrobial compositions comprising any of the compositions disclosed in the present specification comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract). In some aspects, these antimicrobial compositions can be used as preservatives or bactericides. For example, the antimicrobial compositions of the present disclosure can be used to treat or prevent infections, e.g., by S. aureus, P.

aeruginosa, E. coli, Clostridium botulinum, Clostridium difficile, Helicobacter pylori, Candida albicans, Aspergillus niger, Phytophthora ramrum. In some aspects, the compositions disclosed herein comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) can be used in preservative formulations.

[0208] The antimicrobial compositions disclosed herein are particularly effective as preservatives for personal care products. The antimicrobial compositions disclosed herein can be used in, for example, a household (e.g., personal care), industrial, or institutional product. Preferred personal care products include, but are not limited to, shampoos, lotions (e.g., body lotions), conditioners, and soaps. Suitable household products include, but are not limited to, fabric softeners, laundry detergents, and hard surface cleaners. The antimicrobial compositions of the present invention are useful as antimicrobial, fungicidal, and bactericidal agents (such as against allergens, tree and plant fungi, and plant and tree bacteria) and as preservatives in the papermaking, textile, agricultural, and coating industries and in personal care, household, industrial, and institutional products.

[0209] The antimicrobial composition may be incorporated into substrates susceptible to microbial growth to preserve them. For example, the preservative system may be incorporated into or be a personal care product, such as a shampoo, conditioner, cream, lotion (such as body lotion), cosmetic, or a household product, such as a fabric softener, laundry detergent, or hard surface cleaner; or an industrial product, such as paint, coatings, wood, textile, adhesive, sealant, leather, rope, paper, pulp, paper board, sheet rock, ceiling tiles, plastic, fuel, petroleum, oil, rubber working fluid, metal working fluid, starches (such as pet food starch), or mineral slurry, such as a slurry of clay, calcium carbonate, or titanium oxide (Ti02). See, e.g., U.S. Pat. No. US 6251461 and U.S. Publ. No. US20040091558, which are herein incorporated by reference in their entireties.

(iv) Fungicides

[0210] In some aspects, the compositions disclosed in the present specification can be used as fungicides. Accordingly, the present disclosure provides fungicide compositions comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract). Also provided as a method for controlling fungi in an agricultural crop or on ornamental plants comprising applying a treatment comprising a fungicide composition disclosed herein (i.e., any of the compositions comprising a Bulnesia sp. plant extract and a Humulus sp. plant extract provided in the present disclosure). In some aspects, the fungus is powdery mildew or late blight (Phytophthora infestans). See, e.g., U.S. Publ. No. US20050043404), which is herein incorporated by reference in its entirety.

(v) Aphicides

[0211] In some aspects, the compositions disclosed in the present specification can be used to treat of prevent aphid infestations. Accordingly, the present disclosure provides aphicide compositions comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract). Also provided is a method of controlling aphids in an agricultural crop or on ornamental plants comprising applying a treatment comprising an aphicide composition disclosed herein (i.e., any of the compositions comprising & Bulnesia sp. plant extract and a

Humulus sp. plant extract provided in the present disclosure). In some aspects, the aphid is green peach aphid (Myzus persicae). See, e.g., U.S. Pat. No. US8153146, which is herein incorporated by reference in its entirety.

(vi) Allergen control compositions

[0212] In some aspects, the compositions disclosed in the present specification can be used for allergen control. Accordingly, the present disclosure provides anti -allergenic agents comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract). Also provided are methods for allergen control comprising applying a treatment, for example, to a surface such as a fabric, with a Bulnesia sp. plant extract and a Humulus sp. plant extract provided in the present disclosure. See, e.g., U.S. Publ. No. US20050054702, which is herein incorporated by reference in its entirety.

[0213] The present disclosure provides a method of controlling allergens comprising applying to the allergens an anti-allergenic effective amount of at least one anti-allergenic agent comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract). The present disclosure provides a household formulation comprising an anti-allergenic effective amount of a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract).

[0214] The present disclosure also provides a wipe comprising an anti-allergenic

effective amount of a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract).

[0215] The term "allergens" as used herein includes, but is not limited to, all types of allergens (including household allergens), such as dust mites and dust mite allergens

[0216] The term "allergen control" as used herein refers to killing, inhibiting the growth of, immobilizing, denaturing and/or removing allergens from an environment, animal or substrate.

[0217] An "anti-allergenic effective amount" refers to an amount effective to kill, inhibit the growth of, immobilize, or remove allergens from an environment or a substrate. An anti-allergenic effective amount of any of the anti-allergenic agents described herein or mixtures thereof may be determined by methods known in the art. The anti-allergenic agent and formulations of the present invention can be applied to an environment (such as the air in a room), animal, or a substrate by any method known in the art including, but not limited to, spraying, misting, dusting, brushing, wiping, or sprinkling the anti- allergenic agent in the environment or on the substrate.

[0218] The term "substrate" as used herein includes, but is not limited to, animals

including their skin and fur; hard surfaces such as ceiling tiles, air vents, floors, counters, and tables; textile surfaces such as carpets, rugs, mattresses, bedding materials, upholstery, fabric toys, curtains, and window treatments; and other dust gathering surfaces. The anti-allergenic agents of the present invention may be applied, for example, in households, hotels, hospitals, schools, institutions, airplanes, cars, trains, and boats.

(vii) Veterinary, Pest, and Parasite Control Uses

[0219] In some aspects, the compositions disclosed in the present specification can be used as a veterinary medicament for treating parasitic infestation of animals or pest control (e.g., in domestic environments). Accordingly, the present disclosure provides veterinary medicaments comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract). Also provided as methods of use of the veterinary medicaments disclosed herein comprising the administration of a composition comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) to an animal in need thereof.

[0220] The present disclosure also provides compositions for pest and parasite control in animals comprising a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) and a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract), and methods to use such compositions to control populations of pests (e.g., flies, mosquitoes, etc.) or parasites (e.g., ticks or fleas) by, e.g., spraying, misting, dusting, brushing, wiping, or sprinkling the pest or parasite control composition in the environment, on a substrate, or on an animal.

[0221] The compositions of the invention are preferably used for controlling (including preventing) pest and parasitical infestations and infections of animals, preferably warmblooded animals (including humans) and fish. They are, for example, suitable for controlling and preventing infestations and infections in animals, preferably mammals such as cattle, sheep, swine, camels, deer, horses, pigs, poultry, rabbits, goats, dogs and cats, water buffalo, donkeys, fallow deer and reindeer, and also in fur-bearing animals such as mink, chinchilla and raccoon, birds such as hens, geese, turkeys and ducks and fish such as fresh- and salt-water fish such as trout, carp and eels.

[0222] The compositions of the present disclosure can be particularly used for controlling and preventing infestations and infections in companion animals, such as dogs or cats.

[0223] Infestations in warm-blooded animals and fish include, but are not limited to, fleas, ticks, lice, biting lice, nasal bots, keds, biting flies, muscoid flies, flies, myiastic fly larvae, chiggers, mites, gnats, and mosquitoes. The compositions are suitable for systemic and/or non-systemic control of ecto- and/or endoparasites. They are active against all or some stages of development. The compositions are especially useful for controlling ectoparasites.

[0224] The compositions disclosed herein are especially useful for controlling parasites and pests of the following orders and species, respectively: fleas (Siphonaptera), e.g. Ctenocephalides felis, Ctenocephalides canis, Xenopsylla cheopis, Pulex irritans, Tunga penetrans, and Nosopsyllus fasciatus, and icks and parasitic mites (Parasitiformes): ticks (Ixodida), e.g. Ixodes scapularis, Ixodes holocyclus, Ixodes pacificus, Rhiphicephalus sanguineus, Dermacentor andersoni, Dermacentor variabilis, Amblyomma americanum, Ambryomma maculatum, Ornithodorus hermsi, Ornithodorus turicata and parasitic mites (Mesostigmata), e.g. Ornithonyssus bacoti and Dermanyssus gallinae .

[0225] The compositions disclosed herein are also useful for controlling parasites and pests of the following order and speciues: flies, mosquitoes (Diptera), e.g. Aedes aegypti, Aedes albopictus, Aedes vexans, Anastrepha ludens, Anopheles maculipennis, Anopheles crucians, Anopheles albimanus, Anopheles gambiae, Anopheles freeborni, Anopheles leucosphyrus, Anopheles minimus, Anopheles quadrimaculatus, Calliphora vicina, Chrysomya bezziana, Chrysomya hominivorax, Chrysomya macellaria, Chrysops discalis, Chrysops silacea, Chrysops atlanticus, Cochliomyia hominivorax, Cordylobia anthropophaga, Culicoides furens, Culex pipiens, Culex nigripalpus, Culex

quinquefasciatus, Culex tarsalis, Culiseta inornata, Culiseta melanura, Dermatobia hominis, Fannia canicularis, Gasterophilus intestinalis, Glossina morsitans, Glossina palpalis, Glossina fuscipes, Glossina tachinoides, Haematobia irritans, Haplodiplosis equestris, Hippelates spp., Hypoderma lineata, Leptoconops torrens, Lucilia caprina, Lucilia cuprina, Lucilia sericata, Lycoria pectoralis, Mansonia spp. , Musca domestica, Muscina stabulans, Oestrus ovis, Phlebotomus argentipes, Psorophora columbiae, Psorophora discolor, Prosimulium mixtum, Sarcophaga haemorrhoidalis, Sarcophaga spp. , Simulium vittatum, Stomoxys calcitrans, Tabanus bovinus, Tabanus atratus, Tabanus lineola, and Tabanus similis, lice (Phthiraptera), e.g. Pediculus humanus capitis, Pediculus humanus corporis, Pthirus pubis, Haematopinus eurysternus, Haematopinus suis, Linognathus vituli, Bovicola bovis, Menopon gallinae, Menacanthus stramineus, Solenopotes capillatus, Trimenopon spp., Menopon spp., Trinoton spp., Bovicola spp., Werneckiella spp., Lepikentron spp., Trichodectes spp., and Felicola spp., mites Actinedida (Prostigmata) and Acaridida (Astigmata) e.g. Acarapis spp., Cheyletiella spp., Ornithocheyletia spp., Myobia spp., Psorergates spp., Demodex spp., Trombicula spp., Listrophorus spp., Acarus spp., Tyrophagus spp., Caloglyphus spp., Hypodectes spp., Pterolichus spp., Psoroptes spp., Chorioptes spp., Otodectes spp., Sarcoptes spp., Notoedres spp., Knemidocoptes spp., Cytodites spp., and Laminosioptes spp., bugs (Heteropterida), e.g., Cimex lectularius, Cimex hemipterus, Reduvius senilis, Triatoma spp., Rhodnius spp., Panstrongylus spp. and Arilus critatus, Roundworms, Nematoda, Wipeworms and Trichinosis (Trichosyringida), e.g. Trichinellidae (Trichinella spp.), (Trichuridae) Trichuris spp., Capillaria spp., Rhabditida, e.g., Rhabditis spp.,

Strongyloides spp., Helicephalobus spp., Strongylida, e.g., Strongylus spp. , Ancylostoma spp., Necator americanus, Bunostomum spp. (Hookworm), Trichostrongylus spp., Haemonchus contortus, Ostertagia spp., Cooperia spp., Nematodirus spp., Dictyocaulus spp., Cyathostoma spp., Oesophagostomum spp., Stephanurus dentatus, Ollulanus spp., Chabertia spp. , Stephanurus dentatus, Syngamus trachea, Uncinaria spp. , Globocephalus spp. , Metastrongylus spp., Mueller ius capillar is, Protostrongylus spp. , Angiostrongylus spp. , Parelaphostrongylus spp. , Aleurostrongylus abstrusus, and Dioctophyma renale, Intestinal roundworms (Ascaridida), e.g., Ascaris lumbricoides, Ascaris suum, Ascaridia galli, Parascaris equorum, Enterobius vermicularis (Threadworm), Toxocara canis, Toxascaris leonine, Skrjabinema spp., and Oxyuris equi, Camallanida, e.g., Dracunculus medinensis (guinea worm), Spirurida, e.g., Thelazia spp., Wuchereria spp., Brugia spp., Onchocerca spp., Dirofilaria spp., Dipetalonema spp., Setaria spp., Elaeophora spp., Spirocerca lupi, and Habronema spp., Thorny-headed worms (Acanthocephala), e.g., Acanthocephalus spp. , Macracanthorhynchus hirudinaceus and Oncicola spp., Planarians (Plathelminthes), Flukes (Trematoda), e.g., Faciola spp., Fascioloides magna,

Paragonimus spp. , Dicrocoelium spp. , Fasciolopsis buski, Clonorchis sinensis,

Schistosoma spp., Trichobilharzia spp., Alaria alata, Paragonimus spp., and Nanocyetes spp., Cercomeromorpha, in particular Cestoda (Tapeworms), e.g., Diphyllobothrium spp., Tenia spp., Echinococcus spp., Dipylidium caninum, Multiceps spp., Hymenolepis spp., Mesocestoides spp., Vampirolepis spp. , Moniezia spp. , Anoplocephala spp., Sirometra spp. , Anoplocephala spp., and Hymenolepis spp.

In some aspects, the compositions disclosed herein can be used for combating parasitic insects or parasitic arachnids, for controlling mammalian or avian parasitic arachnids, or for controlling mammalian or avian parasitic nematodes. The present disclosure provides a method for controlling parasites in or on an animal, comprising the step of treating the animal with a parasiticidally effective amount of a composition disclosed herein. In a particular aspect, the animal is a mammal or bird. In a particular aspect, the method of controlling a mammalian or avian parasitic arachnid, insect or nematode, is provided the method comprising contacting the arachnid, insect or nematode with an effective amount of a composition disclosed herein, thereby controlling the arachnid, insect or nematode.

[0227] In some aspects, the treating, preferably contacting, occurs while the parasite (e.g., an arachnid, insect or nematode) is in contact with the animal (e.g., a mammalian or avian host). In some aspects, the contacting disrupts a biological function of the parasite, (e.g., an arachnid, insect or nematode). In some aspects, the biological function is selected, e.g., from respiration, neural activity, locomotion, reproduction, or any other physiological activity required for parasite survival. In some aspects, the contacting kills the parasite. In other aspects, the contacting repels the parasite.

[0228] The present disclosure also provides a method for preventing or reducing the

transmission of a parasite borne disease (e.g, a tick borne disease), the method comprising contacting a host organism, i.e., an animal, with a composition disclosed herein, thereby preventing or reducing the transmission of the parasite borne disease.

[0229] The administration of the compositions disclosed herein to animals is carried out directly or in the form of suitable formulations, topically/dermally, orally, or parenterally. In some aspects, compositions comprising Humulus sp. plant extracts and Bulnesia sp. plant extracts can be provided to a host organism infested with parasites, e.g. mammalian or avian parasitic arachnids such as ticks or mites, in a number of convenient

formulations.

[0230] Formulations comprising Humulus sp. plant extracts and Bulnesia sp. plant

extracts can be applied (e.g., sprayed) directly in an area of infestation or they can be bound to a solid support or encapsulated in a time release material. The methods disclosed herein can be carried out by introducing into a parasite, such as an arachnid (e.g., tick or mite), a sufficient amount of a parasiticide, e.g. an arachnicide composition comprising Humulus sp. plant extracts and Bulnesia sp. plant extracts, to impair growth and/or viability of the target parasite and thereby decrease the population of that pest in or on a host. In some aspects, time-release formulations may find use, particularly for applications to animals which are subject to reinfestation. The method of introducing of the subject pesticide into the target pest can be by any method that reduces survival, reproduction, or growth of the parasite, such as an arachnid (e.g., tick or mite). In one aspect, the pesticide is absorbed by the pest or is ingested. The formulations can be used as powders, soaps or detergents for treatment of infestations of animals or humans, including infestations with ticks or mites. In some instances it may be necessary to adjust the treatment formulation so as to reduce any dermatological effects associated with the treatment.

[0231] The compositions are preferably applied topically to the animals in the form of dips, dusts, powders, collars, medallions, sprays, shampoos, spot-on and pour-on formulations and in ointments or oil-in-water or water-in-oil emulsions. In addition, the compositions may be formulated as ear tags for animals, particularly quadrupeds such as cattle and sheep.

[0232] Further, the present disclosure provides various methods of topically

administering such compositions to the skin, hair or feathers of animals, in particular to mammals and birds, for reducing an established infestation and for inhibiting the transmission of parasites, such as ticks or mites, from one host to another, and to prevent the spread of diseases carried by such pests. The present disclosure, therefore, provides wash or topical skin solutions that are useful as "dips" in which the animal can be immersed, or as pour-on or spot-on formulations containing Humulus sp. plant extracts and Bulnesia sp. plant extracts, which are intended to be applied topically to mammals, such as cattle, sheep, pigs, horses, dogs, cats, and the like, and to birds including poultry, such as turkeys, geese, ducks and chickens.

[0233] For oral administration to warm-blooded animals, the compositions disclosed herein can be formulated as animal feeds, animal feed premixes, animal feed

concentrates, pills, solutions, pastes, suspensions, drenches, gels, tablets, boluses and capsules. In addition, the compositions can be administered to the animals in their drinking water. Alternatively, the compositions can be administered to animals parenterally, for example, by intraruminal, intramuscular, intravenous or subcutaneous injection. The compositions can be dispersed or dissolved in a physiologically acceptable carrier for subcutaneous injection. Alternatively, compositions can be formulated into an implant for subcutaneous administration. In addition the compositions can be transdermally administered to animals. For livestock, the process can consist of applying the solution to the animals in pastures and/or before they arrive in pasture or consists of in applying the solution to the animals before they arrive in the "feed lot".

[0234] Because Humulus sp. plant extracts and Bulnesia sp. plant extracts are Generally

Regarded As Safe (GRAS), the compositions disclosed herein can advantageously be used on live stock up to the time of slaughter and may be to treat animals used in milk or egg production without concern that such food products will be contaminated by toxic chemicals.

[0235] In other aspects, parasiticides, such as arachnicides, are prepared in a dusting

composition or as a powder. Dusting compositions are typically prepared by grinding a composition comprising Humulus sp. plant extracts and Bulnesia sp. plant extracts to a fine powder or by spray drying. The skilled artisan can adjust the conditions used in the spray drying process to achieve particles or granules of a size that facilitates delivery to a host or to a parasite, such as a tick, mite, or other arachnid. Desirably, the powder comprises fine particles that coat the host or parasite, e.g. tick or mite, and all of its body parts. The dusting composition can be applied directly to a host.

[0236] Oral solutions are administered directly. Concentrates are administered orally after prior dilution to the use concentration. Oral solutions and concentrates are prepared according to the state of the art. Solutions for use on the skin are trickled on, spread on, rubbed in, sprinkled on or sprayed on. Solutions for use on the skin are prepared according to the state of the art.

[0237] The present disclosure further provides kits for the treatment or prevention of parasite (e.g, tick, mite, or other arachnid) infestation in or on an animal. In one aspect, the kit includes a composition containing an effective amount of a composition comprising Humulus sp. plant extracts and Bulnesia sp. plant extracts in a form suitable for delivery to a host organism. In some aspects, the kit comprises a container, which contains a parasiticide, (e.g., a miticide). Such containers can be boxes, ampoules, bottles, vials, tubes, bags, pouches, blister-packs, or other suitable container forms known in the art. Such containers can be made of plastic, glass, laminated paper, metal foil, or other materials suitable for holding parasiticides, such as miticides. The kit may comprise a dusting powder, shampoo or dip comprising a composition comprising Humulus sp. plant extracts and Bulnesia sp. plant extracts.

8. Articles of manufacture comprising compositions comprising Bulnesia sp. and Humulus sp. plant extracts

[0238] The disclosure also provides articles of manufacture comprising any one of the compositions disclosed herein comprising a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) and a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract). In some aspects, these composition can be used, e.g., as miticides, insecticides, nematicides, etc. In some specific aspects, the Humulus sp. plant extract and the Bulnesia sp. plant extract act synergistically as miticides.

[0239] In a specific aspect, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) used in the articles of manufacture disclosed herein comprise a Humulus lupulus plant extract obtained using supercritical C0₂ and a Bulnesia sarmientoi plant extract obtained using steam distillation. In some aspects, the Humulus lupulus plant extract and the Bulnesia sarmientoi plant extract act synergistically as miticides.

[0240] In a specific aspect, the compositions disclosed herein (e.g., miticide, nematicide, or insecticide compositions) used in the articles of manufacture disclosed herein comprise a Bulnesia sarmientoi essential oil and a Humulus lupulus aqueous extract. In some aspects, the Bulnesia sarmientoi essential oil and the Humulus lupulus aqueous extract act synergistically as miticides.

[0241] In a specific aspect, the compositions (e.g., miticide, nematicide, or insecticide compositions) used in the articles of manufacture disclosed herein comprise

(i) a Bulnesia sarmientoi plant extract comprising guaiol, bulnesol, hanamyol, alpha-guaiene, beta-caryophyllene, beta-guaiene, guaioxide, alpha- bulnesene, elemol, germacrene-B, guaiol, guaiol isomers, eudesm-5-en-l l- ol, beta-eudesmol, 10-epi-gamma-eudesmol, gamma-eudesmol, alpha- eudesmol, 7-epi-alpha-eudesmol, bulnesol isomers, hanamyol isomers, non-characterized sesquiterpene oxides, non-characterized alcohols, or combinations thereof, and

(ii) a Humulus lupulus plant extract comprising iso alpha acids. [0242] In some aspects, the Bulnesia sarmientoi plant extract and the Humulus lupulus plant extract act synergistically as miticides.

[0243] In some aspects, the article of manufacture comprises:

(a) a first container with a composition contained therein, wherein the composition comprises a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract); and,

(b) a second container with a composition contained therein, wherein the composition comprises a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract).

[0244] In a specific aspect, the article of manufacture comprises:

(a) a first container with a composition contained therein, wherein the composition comprises a Humulus lupulus plant extract obtained using supercritical C02; and

(b) a second container with a composition contained therein, wherein the composition comprises a Bulnesia sarmientoi plant extract obtained using steam distillation.

[0245] In a specific aspect, the article of manufacture comprises:

a first container with a composition contained therein, wherein the composition comprises a Bulnesia sarmientoi essential oil and

a second container with a composition contained therein, wherein the composition comprises a Humulus lupulus aqueous extract.

[0246] In a specific aspect, the article of manufacture comprises:

a first container with a composition contained therein, wherein the composition comprises a Bulnesia sarmientoi plant extract comprising guaiol, bulnesol, hanamyol, alpha-guaiene, beta-caryophyllene, beta-guaiene, guaioxide, alpha-bulnesene, elemol, germacrene-B, guaiol, guaiol isomers, eudesm-5-en-l l-ol, beta-eudesmol, 10-epi- gamma-eudesmol, gamma-eudesmol, alpha-eudesmol, 7-epi-alpha-eudesmol, bulnesol isomers, hanamyol isomers, non-characterized sesquiterpene oxides, non-characterized alcohols, or combinations thereof and

a second container with a composition contained therein, wherein the composition comprises a Humulus lupulus plant extract comprising iso alpha acids. [0247] In some aspects, the article of manufacture further comprises a brochure, printed instructions, label, or package insert directing the user (e.g., a distributor or the final user) to combine the contents of the first container and the second container.

[0248] In some aspects, the Humulus sp. plant extract and/or the Bulnesia sp. plant

extract is a concentrate. In some aspects, the article of manufacture further comprises a brochure, printed instructions, label, or package insert directing the user (e.g., a distributor or the final user) to dilute the contents of the container comprising the concentrate.

[0249] In some aspects, the article of manufacture can comprise a Humulus sp. plant extract (e.g., a Humulus lupulus plant extract) comprising purified natural miticide components present in the Humulus sp. plant extract (e.g., alpha acids), miticide components derived from such natural miticide components (e.g., iso alpha acids), their synthetic or semisynthetic counterparts, or combinations thereof.

[0250] In some aspects, the article of manufacture can comprise a Bulnesia sp. plant extract (e.g., a Bulnesia sarmientoi plant extract) comprising purified natural active components (e.g., miticide components) present in t e Bulnesia sp. plant extract, active components (e.g., miticide components) derived from such natural active components, their synthetic or semisynthetic counterparts, or combinations thereof.

[0251] In some aspects, the article of manufacture further comprises a brochure, printed instructions, label, or package insert directing the user (e.g., a distributor or the final user) to combine the contents of the first and second container in the product of manufacture.

[0252] In some aspects, the article of manufacture comprises, for example, bottle(s), vial(s), cartridge(s), sprayer(s), mister(s), fogger(s), diffusor(s), box(es), gloves, spreader(s), sprinkler(s), syringe(s), or any combination thereof. In some aspect, the article of manufacture comprises one or more containers that can be formed from a variety of materials such as glass and/or plastic. In some aspects, the container holds a composition that can be effective for the control, treatment and/or prevention of a pest (e.g, mite) infestation disclosed herein. In some aspects, the container holds at least one active ingredient of the compositions disclosed herein, for example, a biopesticide (e.g., miticide) composition or formulations

[0253] In some aspects, the article of manufacture comprising the compositions disclosed herein further comprises packaging material. In some aspects, the packaging material can contain, for example, plastic, packing peanuts, cardboard, paper, straw, hay, bubble wrap, biodegradable packing material, instructions of use, or any combination thereof. In some aspects, the packaging material contains, for example, printed instructions of use or administration of any of the compositions disclosed herein.

[0254] In some aspects, the printed instructions further comprise a regimen for use, for example, for preventing, combating, exterminating pests (e.g., mites, insects, nematodes, etc.), or any combination thereof. In some aspects, the article of manufacture comprising the compositions disclosed herein further comprises a label. In some aspects, the label refers to use or administration the compositions disclosed herein. In some aspects, the label suggests, for example, a regimen for use, a regimen for preventing, combating, exterminating pests (e.g., mites, insects, nematodes, etc.) or any combination thereof.

[0255] All patents and publications referred to in the present disclosure are expressly incorporated by reference in their entireties.

EXAMPLES

Example 1

Identification of Miticide Compositions from Plant Extracts

and Combinations Thereof

1. Screening

[0256] Having developed and standardized a laboratory method to test plant extracts on the mortality of spider mites (Tetranychus urticae Koch), 57 different plant extracts were evaluated at concentrations from 0 g/L to 10 g/L. TABLE 7 shows the results from the screening corresponding to Humulus lupulus extracts obtained according to the method described in Example 2.

TABLE 7. Effect of Humulus lupulus plant extracts on the mortality of mobile stages of T. urticae

Concentration (g/L)

0.1 0.2 0.5 1 1.5 2 3.5 5 10

Humulus 17.80 40.00 37.80 73.30 80.00 88.90 95.60 100.00 100.00 lupulus

Extract 2. Synergy analysis

[0257] The extract from Humulus lupulus was combined with an extract from Bulnesia sarmientoi. Starting from the concentrations previously tested, an experiment was designed to find which concentrations could result in higher mite mortality. The combinations that were used in this study are shown in TABLE 8. Humulus lupulus extract concentrations ranged from 0 to 1.5 g/L, whereas concentrations of Bulnesia sarmientoi extract ranged from 0 to 5 g/L.

TABLE 8: Synergy study design.

[0258] The results from applying treatments Tl to T 12 on the mortality of the mite T. urticae are shown in TABLE 9. Tests were run in triplicate.

TABLE 9. Effect of the combination of 2 extracts on the mortality of T. urticae

Treatment % mortality (Tetranychus urticae) Average

Test 1 Test 2 Test 3

Tl 86.67% 80.00% 90.00% 85.56%

T2 82.59% 83.33% 90.00% 85.31%

T3 80.00% 83.33% 96.67% 86.67%

T4 66.67% 63.33% 90.00% 73.33%

T5 30.00% 43.33% 53.33% 42.22%

T6 82.96% 83.33% 80.00% 82.10%

T7 100.00% 100.00% 96.67% 98.89%

T8 86.30% 93.33% 100.00% 93.21% T9 100.00% 100.00% 100.00% 100.00%

T10 93.33% 100.00% 100.00% 97.78%

Ti l 89.63% 100.00% 100.00% 96.54%

T12 0.00% 0.00% 3.33% 1.11%

[0259] These results indicated that treatment T7 was the best performer on the mortality of mobile stages of T. urticae.

[0260] The potential presence of a synergistic effect resulting from the combination of the extracts from Humulus lupulus and Bulnesia sarmientoi was evaluated using the internationally recognized methodology proposed by M. C. Berenbaum to evaluate synergism (M. C. BERENBAUM. Synergy, additivism and antagonism in

immunosuppression. Clin. exp. Immunol. (1977) 28, 1-18)

< 1 for synergy

dose of A. dose of B dose of C dose of X

1 for additivism

B. X. > 1 for antagonism.

[0261] For our case: Ae=Humulus lupulus extract 1.5g/L; Dose of A= Humulus lupulus extract 0.75g/L; Be= Bulnesia sarmientoi extract 5g/L; Dose B= Bulnesia sarmientoi extract 1.5g/L. Thus, the equation would be (0.75/1.5)+(1.5/5)=0.8. Since the value obtained is lower than 1, treatment T7, consisting of 0.75 g/L of Humulus lupulus extract and 1.5 g/L of Bulnesia sarmientoi extract has a synergistic effect.

[0262] Other combinations of extracts from Humulus lupulus and Bulnesia sarmientoi have been shown to have synergistic effects (results not shown).

3. Formulation

[0263] Nineteen formulations comprising combinations of extracts from Humulus lupulus and Bulnesia sarmientoi were prepared. In the development of the miticide formulations based on the synergistic blend of active ingredients disclosed above (i.e., a combination of an extract comprising purified iso alpha acids from Humulus lupulus, and an essential oil from Bulnesia sarmientoi), the concentration of each active ingredient varied from 45%) to 80%). Different alcohols were used as solvents including ethanol, isopropyl alcohol, butanol, propanol, and glycerol. Emulsifiers included TWEEN® family products (polysorbate-type nonionic surfactants), SPAN® family products (sorbitan-type nonanionic surfactants), and others. [0264] Carrier vegetal oils were used. Carrier vegetal oils were obtained from different kind of seeds including but not limited to canola, castor, soybean, and sunflower. Mineral oil was also used as a carrier. Long chain fatty acid esters, including octanoyl glycerate, methyl oleate, and others, were also used as carriers.

[0265] The addition of any rheology modifier is recommended, as well as aqueous and non-aqueous dispersants and other coadjuvants.

[0266] One of the big challenges in developing miticide formulations comprising

Bulnesia sarmientoi plants extracts, wherein the extract is an essential oil is that such essential oils are solid at room temperature. Therefore, unless specific formulation conditions are used the formulation are not stable. Accordingly, it is necessary to use conditions in which a natural property of the active ingredient, i.e., solubility, is modified to generate a stable formulation.

[0267] Pilot formulation compositions are shown in TABLE 10.

TABLE 10. Pilot formulations composition

[0268] When the stability of the formulations was evaluated, it was found that

formulations F1-F7 and F 19 were stable. Formulations F8-F18 were found to be unstable.

4. Miticide Efficacy of Formulations

[0269] After the stability study 8 out of the 19 pilot formulation showed good stability, and their efficacy on mites mortality were tested. Each one of the formulations was tested at three different doses: 1 cc/L, 1.5 cc/L and 2.5 cc/L to determine the mortality rate of Tetranychus urticae mites.

[0270] T. urticae was used as a model system for mites belonging to the Tetranychidae family, which includes genera of pest mites such as Tetranychus, Panonichus,

Olygonychus,and Bryobia. The results of the miticide efficacy study are shown in TABLE 11.

TABLE 11. Efficacy of 7 pilot formulations on T. urticae mortality

Formulation Dose Mites mortality

Fl 33.33%

F2 28.86%

F3 75.43%

F4 67.53%

1.0 cc/L

F5 68.86%

F6 36.66%

F7 51.73%

F19 90.00%

Fl 82.20%

F2 82.20%

F3 85.56%

F4 81.10%

1.5 cc/L

F5 72.23%

F6 43.30%

F7 59.96%

F19 90.00% Fl 97.80%

F2 97.80%

F3 95.56%

F4 81.10%

2.5 cc/L

F5 91.13%

F6 76.66%

F7 90.00%

F19 96.66%

[0271] According to the results, the formulation F19 showed the best efficacy to cause mortality on mobile stages of T. urticae. The composition of formulation F 19 is presented in TABLE 12.

TABLE 12: Composition of miticide formulation F 19.

Example 2

Preparation and Molecular Characterization of Humulus lupulus Extracts

[0272] Humulus lupulus extracts used in miticide compositions disclosed herein were extracted utilizing supercritical fluids, with C0₂ food grade under a pressure of 280 bar and temperature of 55°C to reach the levels of supercritical fluid for the carbon dioxide. The plant material used was pelletized fresh leaves to reduce the space in the extraction cabin. [0273] Liquid C0₂ was injected in supercritical conditions. The C0₂ extracted the substances contained in the leaves. The C0₂ solvent, containing the substances extracted from the leaves, was recirculated. Then the extract was led to a chamber at room temperature were the pressure was lowered to 60 bar. Under these conditions the C0₂ became a gas and separated from the extract, resulting in an extract completely free of solvent.

[0274] Using this general method a Humulus lupulus plant extract can be obtained

containing, inter alia, the kind of molecules shown in FIG 1.

[0275] Other Humulus sp. plant extracts can be extracted using this method (e.g., using other plant material), using variants of the method, or any other methods known in the art.

The miticide activity of the extracts, alone or in combination with other extracts can be determined using the methods presented in Example 1.

Example 3

Preparation and Molecular Characterization of Bulnesia sarmientoi Extracts

[0276] The Bulnesia sarmientoi extracts used in the present disclosure were prepared by steam distillation. The wood of the plant was chipped into small pieces between 5 to 10mm, the fragments were placed in the extraction tank, and through a hole in the bottom of the thank a steam flow was injected. Steam passing through the wood at a temperature close to 100°C was capable of melting the compounds and then extracting them. The blend of steam and essential oil passed through a condenser to a temperature of 60°C, making the water and oil vapor condense. Once in liquid form, both components were subjected to a phase separation where at the bottom the water was separated leaving the essential oil pure. The essential oil solidified at room temperature.

[0277] The main components in the B. samientoi essential oil were guaiol, bulnesol, and hanamyol (FIG. 4). Other components identified in the essential oil were alpha-guaiene, beta-caryophyllene, beta-guaiene, guaioxide, alpha-bulnesene, elemol, germacrene-B, guaiol, guaiol isomers, eudesm-5-en-l l-ol, beta-eudesmol, 10-epi-gamma-eudesmol, gamma-eudesmol, alpha-eudesmol, 7-epi-alpha-eudesmol, bulnesol isomers, hanamyol isomers, multiple non-characterized sesquiterpene oxides, and multiple non-characterized alcohols. [0278] Commercial "oil of guaiac" can be used in the miticide compositions disclosed herein and to practice the methods disclosed herein. Oil of guaiac is also produced through steam distillation of a mixture of Bulnesia sarmientoi wood and sawdust. Oil of guaiac is sometimes incorrectly called guaiac wood concrete. It is a yellow to greenish yellow semi-solid mass which melts around 40-50 °C. Once melted, it can be cooled back to room temperature yet remain liquid for a long time. Oil of guaiac is primarily composed of 42-72% guaiol, bulnesol, δ-bulnesene, β-bulnesene, a-guaiene, guaioxide and β-patchoulene. It is considered non-irritating, non-sensitizing, and non-toxic to human skin.

[0279] Other Bulnesia sp. plant extracts can be extracted using this method (e.g., using other plant material), using variants of the method, or any other methods known in the art. The miticide activity of the extracts, alone or in combination with other extracts can be determined using the methods presented in Example 1.

Example 4

Compositions comprising Bulnesia sp. and Humulus sp. plant extracts as

acaricides, miticides, arachnicides, or bactericides

[0280] To assess in vivo the acaricide, miticide, and arachnicide capacity of the

compositions comprising Bulnesia sp. and Humulus sp. plant extracts disclosed herein, their ability to kill, repel infestions, or prevent infestation by acari (e.g., ticks), mites (e.g., mites affecting plants, humans, and animals such as pets or bees), or arachnids (e.g., spiders) will be assayed using methods known in the art. Experimental results will show whether the compositions comprising Bulnesia sp. and Humulus sp. plant extracts disclosed herein are effective against ticks such as deer ticks, mites such as spider mites (affecting plants), Varroa destructor (affecting bees), scabies (affecting humans and other animals), or spiders.

[0281] Additionally, the in vivo bactericide capacity of the compositions comprising

Bulnesia sp. and Humulus sp. plant extracts disclosed herein will be assayed using methods known in the art. Experimental results will show whether the compositions comprising Bulnesia sp. and Humulus sp. plant extracts disclosed herein are effective bactericides. [0282] The capacity of compositions comprising Bulnesia sp. and Humulus sp. plant extract disclosed herein to reduce allergens (in particular mite allergens) and therefore their ability to reduce or ameliorate allergic reactions will also be determined using methods known in the art. Experimental results will show whether the compositions comprising Bulnesia sp. and Humulus sp. plant extracts disclosed herein can effectively reduce allergen levels (e.g., in clothes, bedding, surfaces such as carpet, etc.)

Example 5

Compositions comprising Bulnesia sp. and Humulus sp.

extract as insecticides and nematicides

[0283] To assess in vivo the insecticide and nematicide capacity of the miticide

compositions disclosed herein, their ability to kill, repel infestations, or prevent infestation by nematodes (e.g, Meloidogyne sp., Prtaylenchus sp., Radopholus sp.) or insects (e.g., white flies, trips, aphids, ) will be assayed using methods known in the art. Experimental results will show whether the miticide compositions disclosed herein are effective against nematodes and insects affecting plants or animals, or for domestic uses (e.g., killing or repelling insects or nematodes at homes).

Example 6

Synergistic Effect of compositions comprising Bulnesia sp. and Humulus sp.

extracts as insecticides and nematicides

[0284] Additional experiments were performed to determine whether the synergistic effect of Bulnesia sp. and Humulus sp. in the control mite infestations occurred in other biological models. Accordingly, the compositions disclosed in the present application were used to assess (i) mortality on adults of leaf miners (Liriomyza huidobrensis), (ii) mortality on adults of white flies {Trialeurodes vapor ariorum), and (iii) mortality on J2 of nematodes {Meloidogyne sp.). Several treatments were tested to compare the efficacy of different fractions of Humulus lupulus extract standing alone and in mixes with guaiac wood {Bulnesia sarmientoi) oil. Several pilot formulations were tested (TABLES 13 and 14). TABLE 13 : Compositions used to assess efficacy oiHumulus and Bulnesia extracts on pea leaf miner {Liriomyza huidobrensis) and greenhouse white fly {Trialeurodes vaporariorum).

[0285] The Humulus lupulus extract used in treatments Tl, T2, T3, and T10 was a

fraction rich in iso alpha acids. The Humulus lupulus extract used in treatments T7, T8, T9 and Tl 1 was a fraction rich in beta acids. The Bulnesia extract used in the treatments presented in TABLE 13 was Bulnesia sarmientoi essential oil. The concentration of active principles in treatments T10 and Ti l was equivalent to the concentration of active principle in the Acaricide Formulation control (T12). T12 is a combination of a Humulus lupulus extract enriched in iso alpha acids and Bulnesia sarmientoi essential oil specifically formulated as an acaricide. T3 (Absolute control) refers to a water sample corresponding to the same sample volume used in all the samples.

[0286] The experimental results corresponding to mortality of leafminers {Liriomyza huidobrensis) and whiteflies {Trialeurodes vaporariorum) are presented respectively in FIG. 5 and FIG. 6. The combination of Humulus extract rich in alpha acids and Bulnesia extract was found to have an effect on the mortality of leafminers {Liriomyza

huidobrensis) and whiteflies {Trialeurodes vaporariorum) which was not additive, but synergistic, as evidenced by Berenbaum analysis of the mortality data as explained in Example 1. In both cases, the analysis according to the Berembaum formula yielded values below 1, which are indicative of synergy.

TABLE 14: Compositions used to assess efficacy of Humulus and Bulnesia extracts knot nematode (Meloidogyne sp.)

Treatment Active component Active Component Concentration

Nl Bulnesia oil 0.25 cc/L

N2 Bulnesia oil 0.5 cc/L

N3 Bulnesia oil 1 cc/L

N4 Bulnesia oil 1.5 cc/L

N5 Humulus β extract 0.25 cc/L

N6 Humulus β extract 0.5 cc/L

N7 Humulus β extract 1 cc/L

N8 Humulus β extract 1.5 cc/L

N9 Humulus a extract 0.25 cc/L

N10 Humulus a extract 0.50 cc/L

Ni l Humulus a extract 1 cc/L

N12 Humulus a extract 1.5 cc/L

N13 Humulus β extract + Bulnesia oil 0.375 cc/L + 0.375 cc/L

N14 Humulus β extract + Bulnesia oil 0.6 cc/L + 0.2 cc/L

N15 Humulus β extract + Bulnesia oil 0.75 cc/L + 0.25 cc/L

N16 Humulus β extract + Bulnesia oil 1.125 cc/L + 0.125 cc/L

N17 Humulus β extract formulation 0.8 cc/L

N18 Humulus β extract formulation 1.75 cc/L

N19 Humulus β extract formulation 2.5 cc/L

N20 Acaricide Formulation control 1.4 cc/L

N21 Acaricide Formulation Control 2.5 cc/L

N22 Acaricide Formulation Control 4 cc/L

N23 Acaricide Formulation Control 4.5 cc/L N24 Absolute Control Water only

[0287] The Humulus lupulus extracts used in the treatments presented in TABLE 14 were fractions rich in iso alpha acids ("Humulus a extract") or rich in beta acids ("Humulus β extract"). The Bulnesia extract used in the treatments presented in TABLE 14 was Bulnesia sarmientoi essential oil.

[0288] The concentration of active principles in treatments N13 to T16 was equivalent to the concentration of active principles in the Acaricide Formulation controls (N20 to N23). The Acaricide Formulation control is a combination of a Humulus lupulus extract enriched in iso alpha acids and Bulnesia sarmientoi essential oil specifically formulated as an acaricide. N24 (Absolute control) refers to a water sample corresponding to the same sample volume used in all the samples.

[0289] The experimental results corresponding to mortality of root-knot nematode

(Meloidogyne sp.) are presented in FIG. 7. The combination of Humulus extract rich in alpha acids and Bulnesia extract was found to have an effect on the mortality of root-knot nematode (Meloidogyne sp.) which was not additive, but synergistic, as evidenced by Berenbaum analysis of the mortality data as explained in Example 1. As explained above, analyses according to the Berembaum formula yielding values below 1 are indicative of synergy. The observed synergy was particularly high in the case of nematodes

(Berembaum equation value = 0.13).

***

[0290] It is to be appreciated that the Detailed Description section, and not the Summary and Abstract sections, is intended to be used to interpret the claims. The Summary and Abstract sections sets forth one or more but not all exemplary aspects of the present disclosure as contemplated by the inventor(s), and thus, are not intended to limit the present disclosure and the appended claims in any way.

[0291] The present disclosure has been described above with the aid of functional

building blocks illustrating the implementation of specified functions and relationships thereof. The boundaries of these functional building blocks have been arbitrarily defined herein for the convenience of the description. Alternate boundaries can be defined so long as the specified functions and relationships thereof are appropriately performed. [0292] The foregoing description of the specific aspects will so fully reveal the general nature of the disclosure that others can, by applying knowledge within the skill of the art, readily modify and/or adapt for various applications such specific aspects, without undue experimentation, without departing from the general concept of the present disclosure. Therefore, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed aspects, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance.

[0293] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary aspects, but should be defined only in accordance with the following claims and their equivalents.

Claims

WHAT IS CLAIMED IS:

1. A miticide composition comprising a Bulnesia sp. extract and a Humulus sp. plant extract, wherein the Bulnesia sp. extract and the Humulus sp. plant extract act synergistically as miticides.

2. The miticide composition according to claim 1, wherein the Bulnesia sp. plant is selected from the group consisting of Bulnesia arbor ea, Bulnesia bonariensis, Bulnesia carrapo, Bulnesia chilensis, Bulnesia foliosa, Bulnesia loraniensis, Bulnesia macrocarpa, Bulnesia rivas-martinezii, Bulnesia retama, Bulnesia sarmientoi, Bulnesia schickendantzii, and a combination thereof.

3. The miticide composition according to claim 1, wherein the Bulnesia sp. plant is Bulnesia sarmientoi.

4. The miticide composition according to claim 1, wherein the Humulus sp. plant is selected from the group consisting of Humulus japonicus, Humulus lupulus, Humulus

yunnanensis, and a combination thereof.

5. The miticide composition according to claim 1, wherein the Humulus sp. plant is

Humulus lupulus.

6. The miticide composition according to claim 1, wherein the Humulus sp. plant extract is obtained from Humulus sp. leaves, stems, bark, roots, flowers, buds (hop cones), or any combination thereof.

7. The miticide composition according to claim 6, wherein the Humulus sp. leaves are fresh leaves.

8. The miticide composition according to claim 7, wherein the fresh leaves are pelletized.

9. The miticide composition according to claim 1, wherein the Humulus sp. plant extract is a supercritical C0₂ extract, solvent extract, soxhlet extract, ultrasound extraction extract, or any combination thereof.

10. The miticide composition according to claim 1, wherein the Humulus sp. plant extract consists or consists essentially of a blend of Humulus sp. acids with a high concentration of iso alpha acids.

11. The miticide composition according to claim 10, wherein the concentration of iso alpha acids is at least 75%.

12. The miticide composition according to claim 10, wherein the iso alpha acids are derived from humulone, adhumulone, cohumulone, posthumulone, prehumulone, or combinations thereof.

13. The miticide composition according to claim 1, wherein the Bulnesia sp. plant extract is obtained from Bulnesia sp. wood, bark, roots, stems, leaves, flowers, fruits, seeds, or any combination thereof.

14. The miticide composition according to claim 1, wherein the Bulnesia sp. plant extract is obtained by steam flow extraction.

15. The miticide composition according to claim 1, wherein the Bulnesia sp. plant extract is an essential oil.

16. The miticide composition according to claim 1, wherein the Bulnesia sp. plant extract comprises guaiol, bulnesol, hanamyol, or combinations thereof.

17. The miticide composition according to claim 16, wherein the Bulnesia sp. plant extract further comprises a-guaiene, β-caryophyllene, β-guaiene, guaioxide, a-bulnesene, elemol, germacrene-B, eudesm-5-en-l l-ol, B-eudesmol, lO-epi-y-eudesmol, γ-eudesmol, a-eudesmol, 7-epi-a-eudesmol, or combinations thereof.

18. The miticide composition according to claim 1, wherein the Bulnesia sp. plant extract is an essential oil from Bulnesia sarmientoi wood and the Humulus sp. plant extract comprises Humulus sp. acids from Humulus lupulus leaves with a concentration of iso alpha acids of at least 75%.

19. The miticide composition according to claim 18, wherein the ratio of essential oil from Bulnesia sarmientoi wood and the Humulus sp. plant extract is about 6: 1 to 1 :6.

20. A miticide formulation comprising the miticide composition according to any one of claims 1 to 19 and further comprising at least a solvent, a surfactant, an emulsifier, a carrier, or a combination thereof.

21. The miticide formulation according to claim 20, wherein the surfactant or emulsifier is a cationic, anionic or nonionic surfactant or combinations thereof.

22. The miticide formulation according to claim 20, wherein the surfactant comprises

TWEEN® 85 (polyoxyethylenesorbitan trioleate).

23. The miticide formulation according to claim 20, wherein the carrier is selected from the group consisting of canola oil, castor oil, soybean oil, sunflower oil, mineral oil, long chain fatty acid ester, sesame oil, and combination thereof.

24. The miticide formulation according to claim 23, wherein the long chain fatty acid ester is selected from octanoyl glycerate, methyl oleate, and a combination thereof.

25. The miticide formulation according to claim 23, wherein the carrier comprises canola oil.

26. The miticide formulation according to claim 20, wherein the solvent is selected from the group consisting of ethanol, isopropyl alcohol, butanol, propanol, glycerol, methanol, ethyl lactate, and combinations thereof.

27. The miticide formulation according to claim 25, wherein the solvent comprises ethanol.

28. A method for controlling or preventing mite infestation and/or mite damage to plants, comprising treating the plants or the locus surrounding the plants with a miticidally effective amount of the miticide composition according to any one of claims 1 to 29, or the miticide formulation according to any one claims 20 to 27.

29. A method for controlling or preventing insect infestation and/or insect damage to plants, comprising treating the plants or the locus surrounding the plants with an effective amount of the composition according to any one of claims 1 to 29, or the formulation according to any one claims 20 to 27.

30. A method for controlling or preventing nematode infestation and/or nematode damage to plants, comprising treating the plants or the locus surrounding the plants with an effective amount of the composition according to any one of claims 1 to 29, or the formulation according to any one claims 20 to 27.

31. A method to treat a surface to prevent mite infestation with a miticidally effective amount of the miticide composition according to any one of claims 1 to 29, or the miticide formulation according to any one claims 20 to 27.

32. A method to treat a surface to prevent insect infestation with an effective amount of the composition according to any one of claims 1 to 29, or the formulation according to any one claims 20 to 27.

33. A method to treat a surface to prevent nematode infestation with an effective amount of the composition according to any one of claims 1 to 29, or the formulation according to any one claims 20 to 27.

34. The method according to claim 28 or claim 31, wherein the mite is Tetranychus urticae.

35. The method according to claim 29 or claim 32, wherein the insect is selected from the group consisting of white flies, trips and aphids.

36. The method according to claim 30 or claim 33, wherein the nematode is selected from the group consisting of Meloidogyne sp. , Prtaylenchus sp. , and Radopholus sp..

37. The method according to claims 31 to 33, wherein the surface is in an agricultural, a horticultural, or a garden environment.

38. The method according to any one of claims 28 to 37, further comprising a dose regimen.

39. The method according to claim 39, wherein the dose regimen comprises at least one daily dose.

40. The method according to claim 38, wherein the dose regimen comprises at least one weekly dose.