Antimicrobial extract from vibrio gazogenes and methods of producing and using the same

Abstract

This invention relates to an antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943 and methods of producing and using the same to inhibit microbial growth and microbial infections of plants and animals.

Description

Attorney Docket No.1655.3.WO ANTIMICROBIAL EXTRACT FROM VIBRIO GAZOGENES AND METHODS OF PRODUCING AND USING THE SAME STATEMENT OF PRIORITY

[0001] This application claims the benefit of U.S. Provisional Application Serial No.63 / 582,527, filed September 14, 2023, the entire contents of which are incorporated by reference herein. FIELD OF THE INVENTION

[0002] This invention relates to an antimicrobial extract from Vibrio gazogenes, ATCC Accession No. 43943, and methods for producing and using the same to inhibit the growth of bacterial and fungal pathogens and plants and animals. BACKGROUND OF THE INVENTION

[0003] Pathogenic bacteria and fungi pose a significant threat to human health and agriculture. There is a constant need for new and effective antimicrobial agents that can combat these pathogens. While many synthetic antimicrobials have been developed over the years, they often suffer from problems such as toxicity, resistance, and environmental persistence. Therefore, there is a need for the development of novel antimicrobial agents that are effective against a broad range of microorganisms.

[0004] Vibrio gazogenes is a gram-negative, rod-shaped bacterium that has been isolated from various marine environments. While this bacterium is not known to cause human infections, it has been found to produce a variety of bioactive compounds and one of the most well-known natural products that it makes is prodigiosin, which has antimicrobial properties. Extracts of live and heat inactivated Vibrio gazogenes, ATCC Accession No.43942 have been shown to inhibit aflatoxin production in Aspergillus flavus dose-dependently. See US 2022 / 0249576 A1. However, such extracts have not been shown to exhibit antimicrobial activity. SUMMARY OF THE INVENTION

[0005] One aspect of the invention provides a method of producing an antimicrobial extract comprising (a) contacting cells of Vibrio gazogenes, ATCC Accession No.43943, with methanol; and (b) extracting the cells of (a) with one or more of acetone, chloroform, acetonitrile, orAttorney Docket No.1655.3.WO dimethylsulfoxide, thereby producing the antimicrobial extract, the extract being substantially devoid of membrane components and comprising compounds soluble in (i) methanol and (ii) acetone, chloroform, acetonitrile, isopropanol, and / or dimethylsulfoxide, thereby producing an antimicrobial extract.

[0006] In a second aspect, the invention provides a method for preventing or reducing the growth of a microbial pathogen comprising contacting a microbial pathogen with an effective amount of an antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, the extract being substantially devoid of membrane components and comprising compounds soluble in (a) methanol and (b) acetone, ethanol, chloroform, acetonitrile, isopropanol, or dimethylsulfoxide, thereby preventing or reducing the growth of a microbial pathogen.

[0007] In a third aspect of the invention is provided a method for preventing or reducing the likelihood of contamination of a food product by a microbial pathogen comprising applying to the food product an effective amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, the extract being substantially devoid of membrane components and comprising compounds soluble in (a) methanol and (b) acetone, ethanol, chloroform, acetonitrile, isopropanol, or dimethylsulfoxide, thereby preventing or reducing the likelihood of contamination of a food product by a microbial pathogen.

[0008] In a fourth aspect, a method for inhibiting the growth of a microbial pathogen on an inanimate surface or environmental sample is provided, the method comprising contacting an inanimate surface or environmental sample with an effective amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, the extract being substantially devoid of membrane components and comprising compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide, thereby inhibiting the growth of a microbial pathogen on an inanimate surface or environmental sample.

[0009] In a fifth aspect, the invention provides a method for preventing, treating, or reducing the likelihood of infection by a microbial pathogen comprising administering to a subject in need thereof an effective amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, the extract being substantially devoid of membrane components and comprising compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide, thereby preventing, treating, or reducing the likelihood of infection by a microbial pathogen.Attorney Docket No.1655.3.WO

[0010] In a six aspect, the invention provides a method of treating, preventing or reducing the likelihood of infection of a plant or part thereof by a microbial pathogen comprising contacting the plant or part thereof with an effective amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, the extract being substantially devoid of membrane components and comprising compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide, thereby treating, preventing or reducing the likelihood of infection of a plant or part thereof by a microbial pathogen.

[0011] In a further aspect, the invention provides an antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, the extract being substantially devoid of membrane components and comprising compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide.

[0012] These and other aspects of the invention are set forth in more detail in the description of the invention below. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIG. 1 shows the antimicrobial activity of extracts from Vibrio gazogenes, ATCC Accession No. 43943, as compared to Vibrio gazogenes, ATCC Accession Nos. 43939, 43940, 43941, 43942, and 43944, as determined by measuring colony diameter of Aspergillus flavus colonies on agar after 3 days.

[0014] FIG. 2 shows LC-MS / MS analysis of cellular extracts from Vibrio gazogenes, ATCC Accession Nos.43939 to 43944.

[0015] FIG. 3 shows the absence of antimicrobial activity of a Vibrio gazogenes, ATCC Accession No. 43942 extract against Aspergillus flavus when applied at 0-, 0.25-, 1- and 3-fold dose relative to the dose used for a Vibrio gazogenes, ATCC Accession No.43943 extract.

[0016] FIG. 4 shows spectrophotometric analysis of two independent extracts of Vibrio gazogenes, ATCC 43943, as compared to pure Prodigiosin.

[0017] FIG. 5 shows antimicrobial activity of Vibrio gazogenes, ATCC Accession No. 43943 against agriculturally relevant fungal pathogens Rhizoctonia solani, Penicillium expansum, Botrytis cinerea, Verticillium dahlia, Fusarium oxysporum, and Phoma betae.

[0018] FIGS. 6A-6C show that treatment of corn seed with an extract of Vibrio gazogenes, ATCC Accession No.43943, promoted the overall health and growth rate of corn seedlings. FIG.Attorney Docket No.1655.3.WO 6A, Growth rate at growth stages (V2 to V5) by treatment. FIG. 6B, Stalk diameter at V6 by treatment. W, average widest stalk diameter; N, average narrowest stalk diameter. Error bars indicate standard deviation. FIG. 6C, Plant dry weight at V7 by treatment. Error bars indicate standard deviation. Statistical significance of differences among the treatments for each measured parameter (growth stages, stalk diameters, and plant dry weight) was determined using one-way ANOVA (Analysis of Variance) with n=4. A p-value <0.05 was deemed as a significant difference. Comparisons were made with the control group (No Inoculant); ** denotes p-value less than 0.0001. DETAILED DESCRIPTION

[0019] The present invention now will be described hereinafter with reference to the accompanying drawings and examples, in which embodiments of the invention are shown. This description is not intended to be a detailed catalog of all the different ways in which the invention may be implemented, or all the features that may be added to the instant invention. For example, features illustrated with respect to one embodiment may be incorporated into other embodiments, and features illustrated with respect to a particular embodiment may be deleted from that embodiment. Thus, the invention contemplates that in some embodiments of the invention, any feature or combination of features set forth herein can be excluded or omitted. In addition, numerous variations and additions to the various embodiments suggested herein will be apparent to those skilled in the art in light of the instant disclosure, which do not depart from the instant invention. Hence, the following descriptions are intended to illustrate some particular embodiments of the invention, and not to exhaustively specify all permutations, combinations and variations thereof.

[0020] Unless otherwise defined, 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 invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0021] All publications, patent applications, patents and other references cited herein are incorporated by reference in their entireties for the teachings relevant to the sentence and / or paragraph in which the reference is presented.Attorney Docket No.1655.3.WO

[0022] Unless the context indicates otherwise, it is specifically intended that the various features of the invention described herein can be used in any combination. Moreover, the present invention also contemplates that in some embodiments of the invention, any feature or combination of features set forth herein can be excluded or omitted. To illustrate, if the specification states that a composition comprises components A, B and C, it is specifically intended that any of A, B or C, or a combination thereof, can be omitted and disclaimed singularly or in any combination.

[0023] As used in the description of the invention and the appended claims, the singular forms "a," "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0024] Also as used herein, "and / or" refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative ("or").

[0025] The term "about," as used herein when referring to a measurable value such as an amount or concentration and the like, is meant to encompass variations of ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of the specified value as well as the specified value. For example, "about X" where X is the measurable value, is meant to include X as well as variations of ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of X. A range provided herein for a measureable value may include any other range and / or individual value therein.

[0026] As used herein, phrases such as "between X and Y" and "between about X and Y" should be interpreted to include X and Y. As used herein, phrases such as "between about X and Y" mean "between about X and about Y" and phrases such as "from about X to Y" mean "from about X to about Y."

[0027] Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. For example, if the range 10 to 15 is disclosed, then 11, 12, 13, and 14 are also disclosed.

[0028] The term "comprise," "comprises" and "comprising" as used herein, specify the presence of the stated features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof.Attorney Docket No.1655.3.WO

[0029] As used herein, the transitional phrase "consisting essentially of" means that the scope of a claim is to be interpreted to encompass the specified materials or steps recited in the claim and those that do not materially affect the basic and novel characteristic(s) of the claimed invention. Thus, the term "consisting essentially of" when used in a claim of this invention is not intended to be interpreted to be equivalent to "comprising."

[0030] As used herein, the terms "reduce," "reduced," "reducing," "reduction," "inhibiting,” “inhibit,” “preventing” and "decrease" (and grammatical variations thereof), describe, for example, a decrease of at least about 5%, 10%, 15%, 20%, 25%, 35%, 50%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100% as compared to a control. For example, a microbial pathogen contacted with an antimicrobial extract as described herein can exhibit a decrease in growth that is at least about 50% or greater than that of a control microbial pathogen not contacted with the antimicrobial extract. In some aspects, the growth of a microbial pathogen is inhibited by between 50% and 90% as compared to a control microbial pathogen not contacted with the antimicrobial extract. In some embodiments, a comparison with a control is made under the same growth conditions, e.g., the same environmental conditions (growth medium, hydration, light, heat, nutrients and the like). In some embodiments, the reduction can result in no or essentially no (i.e., an insignificant amount, e.g., less than about 10% or even 5%) microbial growth.

[0031] As used herein, the terms "increase," "increasing," "increased," "enhance," "enhanced," "enhancing," and "enhancement" (and grammatical variations thereof) describe an elevation of at least about 5%, 10%, 15%, 20%, 25%, 50%, 75%, 100%, 150%, 200%, 300%, 400%, 500% or more as compared to a control.

[0032] The microorganisms and particular strain described herein, unless specifically noted otherwise, are all separated from nature and grown under artificial conditions such as in shake flask cultures or through scaled-up manufacturing processes, such as in bioreactors to maximize bioactive metabolite production, for example. Growth under such conditions leads to strain “domestication.” Generally, such a “domesticated” strain differs from its counterparts found in nature in that it is cultured as a homogenous population that is not subject to the selection pressures found in the natural environment but rather to artificial selection pressures.

[0033] As used herein, the term “biologically pure isolate” refers to a microbial culture that is free or essentially free of biological contamination and that has genetic uniformity such thatAttorney Docket No.1655.3.WO different subcultures taken therefrom will exhibit identical or substantially identical genotypes and phenotypes. In some embodiments, the biologically pure culture is 100% pure (i.e., all subcultures taken therefrom exhibit identical genotypes and phenotypes). In some embodiments, the biologically pure culture is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, 99.6, 99.7, 99.8, or 99.9% pure (i.e., at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, 99.6, 99.7, 99.8, or 99.9% of the subcultures taken therefrom exhibit identical genotypes and phenotypes).

[0034] As used herein, the terms “inoculant” and “inoculum” refer to a composition comprising microbial cells, wherein the cells are capable of propagating on or in a suitable growth medium or substrate (e.g., a soil) when conditions (e.g., temperature, moisture, nutrient availability, pH, etc.) are favorable for microbial growth.

[0035] As used herein, the terms “colony forming unit” and “cfu” refer to a unit which estimates the number of microbial cells in a sample that are viable and able to multiply in a suitable growth medium or substrate (e.g., a soil) when conditions (e.g., temperature, moisture, nutrient availability, pH, etc.) are favorable for microbial growth.

[0036] As used herein, the term “pathogen” takes on the ordinary and customary meaning of microbes that cause infection in a host. Within the scope, pathogens of interest include, but are not limited to, those that cause human and / or plant infections, and these generally comprise bacteria and fungi. Further, pathogens recognized as microbes that cause infection by being transferred indirectly through contaminated surfaces are of interest within the scope of the disclosure.

[0037] The term "MIC" or "minimal inhibitory concentration" refers to the lowest concentration of an antibiotic that is necessary to inhibit the growth of a bacterial isolate in vitro. A common method for determining the MIC of an antibiotic is to prepare several tubes with serial dilutions of the antibiotic, which are then inoculated with the bacterial isolate of interest. The MIC of an antibiotic can be determined from the tube with the lowest concentration which does not show any turbidity (no growth).

[0038] "IC50" refers to an amount, concentration or dosage of a particular test compound that achieves a 50% inhibition of a maximal response, such as cell growth or proliferation measured via an in vitro or cell-based assay.

[0039] As used herein, the term “effective amount,” “effective concentration” or “effective amount / concentration” refers to an amount or concentration that is sufficient to cause a desired effect (e.g., inhibiting growth of a microbe). The absolute value of the amount / concentration thatAttorney Docket No.1655.3.WO is sufficient to cause the desired effect may be affected by factors such as the type and magnitude of effect desired, the type, size, and volume of material to which the antimicrobial extract will be applied, the stability of the antimicrobial extract and the storage conditions (e.g., temperature, relative humidity, duration). Those skilled in the art will understand how to select an effective amount / concentration using routine dose-response experiments.

[0040] As used herein, the terms “treat,” “treatment,” and “treating” refer to the reduction or amelioration of the progression, severity, and / or duration of a microbial infection, or the amelioration of one or more symptoms (preferably, one or more discernible symptoms) of a microbial infection resulting from the administration of an antimicrobial extract of the invention. In some embodiments, the terms “treat,” “treatment,” and “treating” refer to the amelioration of at least one measurable physical parameter of a microbial infection. In some embodiments, the terms “treat,” “treatment,” and “treating” refer to the inhibition of the progression of a microbial infection, either physically by, e.g., stabilization of a discernible symptom, physiologically by, e.g., stabilization of a physical parameter, or both. The terms “treat,” “treatment,” and “treating” may also refer to the reduction in the amount of a pathogenic microorganism on / in an inanimate surface or environmental sample.

[0041] As used herein, the terms "prevent," "preventing" or "prevention" refer to any type of action that results in the absence, avoidance and / or delay of the onset and / or progression of a disease, disorder and / or a clinical symptom(s) in a subject or a plant and / or a reduction in the severity of the onset of the disease, disorder and / or clinical symptom(s) relative to what would occur in the absence of the methods of the invention. The prevention can be complete, e.g., the total absence of the disease, disorder and / or clinical symptom(s). The prevention can also be partial, such that the occurrence of the disease, disorder and / or clinical symptom(s) in the subject or plant and / or the severity of onset is less than what would occur in the absence of the present invention.

[0042] The term “reducing the likelihood” refers to the fact that in any given population of patients, subjects, plants or samples, the antimicrobial extract of the invention may be used to reduce the likelihood of an occurrence or recurrence of infection by or contamination with a microbial pathogen in one or more subjects, plants, or samples.

[0043] As used herein, the terms “subject,” “patient,” and “animal” may be used interchangeably and may refer to, for example, a bird, a mammal, a reptile, or a fish. In some embodiments, aAttorney Docket No.1655.3.WO mammal may include, but is not limited to, a non-primate (e.g., a cow, pig, horse, sheep, goat, rabbit, guinea pig, rat, cat, dog, or mouse) or a primate (e.g., a monkey, chimpanzee, gorilla, or a human). In some embodiments, the mammal may be a human. In some embodiments, the subject may be a non-human animal such as a farm animal (e.g., a horse, cow, pig, or sheep), or a pet (e.g., a dog, cat, guinea pig, or rabbit). In some embodiments, the subject is a human.

[0044] The present disclosure relates to an antimicrobial extract obtained from cells of Vibrio gazogenes, ATCC (American Type Culture Collection, Manassas, VA) Accession No.43943, and use of the same in methods for preventing or reducing the growth of a microbial pathogen; preventing or reducing the likelihood of contamination of a food product by a microbial pathogen; inhibiting the growth of a microbial pathogen on an inanimate surface or environmental sample; preventing, treating, or reducing the likelihood of infection of a subject by a microbial pathogen; and / or treating, preventing or reducing the likelihood of infection of a plant or part thereof by a microbial pathogen. In addition, the present disclosure provides for the preparation of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, by (a) contacting cells of Vibrio gazogenes, ATCC Accession No.43943, with methanol; and (b) extracting the cells of (a) with one or more of acetone, chloroform, acetonitrile, isopropanol, or dimethylsulfoxide, thereby producing the antimicrobial extract, wherein the extract is substantially devoid of membrane components and comprises compounds soluble in (i) methanol and (ii) acetone, chloroform, acetonitrile, isopropanol, and / or dimethylsulfoxide.

[0045] As used herein, “extract” refers to a substance obtained by extracting a raw material with one or more solvents. The term “antimicrobial extract of Vibrio gazogenes” refers to a substance obtained by extracting Vibrio gazogenes cells, in particular Vibrio gazogenes cells of ATCC Accession No. 43943, with one or more solvents, wherein the extracted substance exhibits antimicrobial activity, e.g., antibacterial and / or antifungal activity. In some embodiments, the antimicrobial extract of the invention is substantially devoid or substantially free of membrane components, e.g., membrane lipids and / or membrane proteins. By saying that a antimicrobial extract is “substantially devoid of” or “substantially free of” membrane components, it is meant in the context of the present invention that the antimicrobial extract should not comprise any significant amount of membrane components including but not limited to membrane lipids and / or membrane-localized proteins. A non-significant amount is defined as less than about 15% byAttorney Docket No.1655.3.WO weight, e.g., less than about 10% by weight, less than about 5% by weight, or less than about 1% by weight, based on the dry weight of the antimicrobial extract, of membrane components.

[0046] In some aspects, the antimicrobial extract of the invention is prepared by first contacting cells of Vibrio gazogenes, ATCC Accession No.43943, with a sufficient amount of methanol for a sufficient amount of time to, e.g., kill the bacterium and / or disrupt the excellular membrane. In some aspects, the cells are contacted with methanol for about 30 minutes with shaking. In some aspects, the cells are contacted with methanol for about 30 minutes with shaking at 100 rpm at 25°C. In some aspects, the methanol is 100% methanol. In other aspects, the amount of methanol may be less than 100%, e.g., about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. In some aspects, the cells are suspended in culture medium and the methanol is added to the suspended cells. In other aspects, the cells are pelleted (e.g., centrifuged and the culture medium is removed) and the methanol is added to the pelleted cells. In some aspects, about 1 mL of methanol is added to about 100 µL to about 300 µL of pelleted cells, e.g., the ratio of methanol to cell pellet (v / v) is in the range of about 10:1 to about 10:3, e.g., about 10:1, 10:1.5, 10:2, 10:2.5, or 10:3.

[0047] Subsequent to treatment with methanol, the cells / cellular contents are extracted with a second solvent comprising one or more of acetone, chloroform, acetonitrile, isopropanol, propylene glycol, and / or dimethylsulfoxide. In some aspects, the second solvent is composed of 100% acetone, 100% chloroform, 100% acetonitrile, 100% isopropanol, 100% propylene glycol, or 100% dimethylsulfoxide. In other aspects, the second solvent may be a combination of any one of acetone, chloroform, acetonitrile, isopropanol, propylene glycol, and dimethylsulfoxide, wherein each of the solvents of the combination may be in the range of 1% to 99% (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%), by volume of the total volume of the second solvent. In some embodiments, a combination of 80% acetone and 20% dimethylsulfoxide may be used as the second solvent. In some embodiments, a combination of 60% isopropanol and 40% propylene glycol may used as the second solvent. The amount of second solvent added to the cells / cellular contents may be in a ratio in the range of about 9:1 to about 4:1, e.g., about 9:1, 8:1, 7:1, 6:1, 5:1, or 4:1 second solvent to methanol, e.g., about 5 mL acetone or isopropanol to about 1 mL methanol, or about 5 mL isopropanol to about 1 mL methanol.

[0048] After the addition of methanol and / or extraction with the second solvent, the cells / cellular contents may be centrifuged, filtered or decanted to remove debris or insoluble material. TheAttorney Docket No.1655.3.WO resulting extract of the method of the invention is an antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, that is substantially devoid of membrane components and comprises compounds soluble in (i) methanol and (ii) one or more of acetone, chloroform, acetonitrile, isopropanol, and / or dimethylsulfoxide. A compound that is “soluble” in a solvent refers to the ability of compound, the solute, to form a solution with another substance, the solvent. The extent of the solubility of a compound in a specific solvent is generally measured as the concentration of the solute in a saturated solution, one in which no more solute can be dissolved. More specifically, in accordance with one aspect, the present invention provides an antimicrobial extract comprising compounds that are at least sparingly soluble or soluble in a non-aqueous solvent such as methanol, acetone, ethanol, isopropanol, chloroform, acetonitrile, and / or dimethylsulfoxide. By “sparingly soluble” it is meant that a compound is generally soluble in one of the solvents, and from about 30 and 100 parts by volume of solvent may be used to solubilize the compound. By “soluble” it is meant that about 10 and 30 parts by volume of solvent may be used to solubilize the compound in question.

[0049] In some embodiments, the antimicrobial extract produced as described herein from V. gazogenes, ATCC Accession No. 43943, comprises prodigiosin (4-Methoxy-5-[(5-methyl-4- pentyl-2H-pyrrol-2-ylidene)methyl]-2,2′-bi[1H-pyrrole]; CAS No.: 82-89-3), 6-hydroxycaproic acid (CAS No.: 1191-25-9), acetophenone (1-phenylethanone; CAS No.: 98-86-2), 2'- deoxyadenosine (CAS No.: 958-09-8), baquiloprim (CAS No.: 102280-35-3), and peptides. In some embodiments, an antimicrobial extract produced from V. gazogenes, ATCC Accession No. 43943, comprises, consists essentially of, or consists of between about 30% to about 32% prodigiosin (e.g., about 30%, 30.5%, 31%, 31.5% or 32% prodigiosin), about 3% to about 4% 6- hydroxycaproic acid (e.g., about 3%, 3.5%, or 4% 6-hydroxycaproic acid), about 1% to about 2% acetophenone (e.g., about 1%, 1.5%, or 2% acetophenone), about 0.5% to about 1% 2'- deoxyadenosine (e.g., about 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1% 2'-deoxyadenosine), about 0.01% to about 0.05% baquiloprim (about 0.01%, 0.02%, 0.03%, 0.04%, or 0.05% baquiloprim), and about 5% to about 30% peptides (about 5%, 10%, 15%, 20%, 25%, or 30% peptides), in particular peptides of proteins involved in protein synthesis, wherein said percent is percent distribution of the total metabolites. As used herein, “peptides” refers to portions of full length proteins, e.g., fragments of proteins, such as peptides of about 7 to about 44 amino acids in length. In some embodiments, the antimicrobial extract produced as described herein from V. gazogenes,Attorney Docket No.1655.3.WO ATCC Accession No.43943 includes at least about 7000 distinct peptides, e.g., about 7000, 7100, 7200, 7300, or 7400 distinct peptides, having a mean peptide size of about 14 amino acid residues. In some embodiments, the antimicrobial extract produced as described herein from V. gazogenes, ATCC Accession No.43943, is red.

[0050] The Vibrio gazogenes used herein may be a biologically pure isolate of Vibrio gazogenes, ATCC Accession No. 43943, that has been optionally cultivated in a suitable culture medium to obtain a sufficient biomass for preparing the antimicrobial extract. The Vibrio gazogenes of the present disclosure may be cultured using any suitable method(s), including, but not limited to, liquid-state fermentation and solid-state fermentation. A biologically pure isolate of Vibrio gazogenes, ATCC Accession No.43943, may be obtained from a frozen stock culture or a single colony from an agar plate. In some aspects, the biologically pure isolate of Vibrio gazogenes, ATCC Accession No.43943, is red in color. In some aspects, a biologically pure isolate of Vibrio gazogenes may be used as an inoculant in a liquid culture medium, e.g., marine broth medium; and the cells may be cultivated at a temperarture in the range of about 24°C to about 29°C (e.g., about 24°C, 25°C, 26°C, 27°C, 28°C or 29°C, in particular about 26°C), e.g., in an orbital shaker, for approximately 18 to 24 hours. In some aspects, a biologically pure isolate of Vibrio gazogene is inoculated into about 5 mL of liquid culture medium, e.g., marine broth medium; the cells are grown at about 26°C for approximately 18 to 24 hours (e.g., about 18, 18.5, 19, 1.5, 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5 or 24 hours or any value or range therein); a portion of the culture (e.g., 1 mL) is transferred to fresh culture medium, e.g., fresh marine broth medium; and the cells are cultivated for approximately an additional 18 to 24 hours. In some aspects, the Vibrio gazogenes cells are cultivated in a volume of about 50 mL to about 200 mL of fresh culture medium, e.g., 75 mL, 100 mL or 150 mL. In some aspects, the Vibrio gazogenes cells are cultivated at about 26°C until an OD600 of about 1.2 is reached. In some aspects, the Vibrio gazogenes cells are cultivated, a small sample (e.g., 1.5 mL) of the culture is removed, the sample is centrifuged to obtain a cellular pellet, the cellular pellet is resuspended in methanol (e.g., about 500 ml) and vortexed, and the resulting extract is analyzed by spectrophotometry (e.g, UV-Visible spectrophotometry) to determine whether the culture has peaks at 535 nm corresponding to prodigiosin as well as two prominent peaks at 465 nm and 505 nm. After cultivation, the cells of the culture may separated from the medium, e.g., by centrifugation, and used in the production of the antimicrobial extract.Attorney Docket No.1655.3.WO

[0051] This invention also provides an antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, produced by the method of the invention. In some aspects, the antimicrobial extract produced by the method of the invention is used as is or may be concentrated by removing all or a portion of the solvents. Thus, in some aspects, the method includes the further step of evaporating all or portion of the methanol and of the one or more of acetone, chloroform, acetonitrile, isopropanol, and / or dimethylsulfoxide to produce a concentrated or dried antimicrobial extract. A concentrated antimicrobial extract will have a lower concentration of solvent and higher relative concentration of solvent soluble components form Vibrio gazogenes than an original or starting extract. A concentrated antimicrobial extract may contain about 1 wt %, 2 wt%, 3 wt%, 4 wt%, 5 wt %, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt %, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt %, 21 wt%, 22 wt%, 23 wt%, 24 wt%, 25 wt%, 26 wt%, 27 wt%, 28 wt%, 29 wt%, 30 wt %, 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35 wt%, 36 wt%, 37 wt%, 38 wt%, 39 wt%, 40 wt %, 41 wt%, 42 wt%, 43 wt%, 44 wt%, 45 wt%, 56 wt%, 57 wt%, 48 wt%, 49 wt%, or <50 wt % of solvent contained in the original extract. This range includes any intermediate value.

[0052] An anhydrous, dry or desiccated antimicrobial extract will have substantially all of the solvent removed from it, for example, an anhydrous, dry or desiccated antimicrobial extract may contain no more than 0.1 wt %, 0.2 wt%, 0.3 wt%, 0.4 wt%, 0.5 wt %, 0.6 wt%, 0.7 wt%, 0.8 wt%, 0.9 wt%, 1 wt %, 2 wt%, 3 wt%, 4 wt%, 5 wt %, 6 wt%, 7 wt%, 8 wt%, 9 wt%, or 10 wt % or any value or range therein of the one or more solvents in the original extract.

[0053] Concentration and dehydration methods are known in the art and include, but are not limited to, those performed with a device such as hot air blower, dryer, cylindrical dryer, zeolite dryer, a desiccator, or a freeze-dryer. An antimicrobial extract may be concentrated or dried by evaporating the solvent using a falling film device, low pressure evaporator, or spray tower. In some aspects, the anhydrous, dry or desiccated antimicrobial extract may be resuspended in one or more of methanol, ethanol, isopropanol, chloroform, acetonitrile, ether, and / or dimethylsulfoxide.

[0054] The antimicrobial extract of the invention may be standardized by removal of the solvent from components extracted from Vibrio gazogenes, weighing the residuum (non-solvent components that dissolved in or were retained in the solvent used for extraction), and admixing, dissolving or suspending the residuum to a desired concentration in a composition. For example,Attorney Docket No.1655.3.WO a known weight of the residuum may be admixed with a known weight of a solid excipient or a known weight of residuum may be dissolved in a solvent, optionally in the presence of a surfactant, stabilizer, buffer or other excipient. The final concentration of the residuum may range from at least 0.001 wt %, 0.005 wt %, 0.01 wt %, 0.05 wt %, 0.1 wt %, 0.5 wt %, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, 21 wt%, 22 wt%, 23 wt%, 24 wt%, 25 wt%, 26 wt%, 27 wt%, 28 wt%, 29 wt%, 30 wt%, 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35 wt%, 36 wt%, 37 wt%, 38 wt%, 39 wt%, 40 wt%, 41 wt%, 42 wt%, 43 wt%, 44 wt%, 45 wt%, 46 wt%, 47 wt%, 48 wt%, 49 wt%, 50 wt%, 51 wt%, 52 wt%, 53 wt%, 54 wt%, 55 wt%, 56 wt%, 57 wt%, 58 wt%, 59 wt%, 60 wt%, 61 wt%, 62 wt%, 63 wt%, 64 wt%, 65 wt%, 66 wt%, 67 wt%, 68 wt%, 69 wt%, 70 wt%, 71 wt%, 72 wt%, 73 wt%, 74 wt%, 75 wt%, 76 wt%, 77 wt%, 78 wt%, 79 wt%, 80 wt%, 81 wt%, 82 wt%, 83 wt%, 84 wt%, 85 wt%, 86 wt%, 87 wt%, 88 wt%, 89 wt%, 90 wt%, 91 wt%, 92 wt%, 93 wt%, 94 wt%, 95 wt%, 96 wt%, 97 wt%, 98 wt%, 99 wt%, or 100 wt % based on the combined weight of the residuum and other components in a final composition. In some aspects, any solid components in a raw antimicrobial extract will be removed by centrifugation or filtration, e.g., through a 0.45 micron or 0.22 micron filter which can also serve to remove microorganisms to produce a purified or sterile solution.

[0055] The activity of the antimicrobial extract may also be standardized based on a quantifiable degree of antimicrobial activity, or other biological activity. For microbes, including gram-positive bacteria, gram-negative bacteria, and fungi, these include standardization based on minimal inhibitory concentration or on the size of a zone of inhibition produced by a known amount or concentration of an antimicrobial extract on a test or control bacterial strain, such as on a Staphylococcus aureus or Aspergillus flavus strain obtained from a depository such as the ATCC. Activity on non-microbial microorganisms, such as viruses, parasites (e.g., protozoa such Plasmodium spp., Entamoeba histolytica, Leishmania spp., and Trypanosoma spp.), or neoplasms may also be determined using conventional assays as can other functional activities of an antimicrobial extract such as on promotion of wound or tissue healing.

[0056] Whether used as is, dried, or dried and resuspended in one or a combination of solvents, the antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, may be used alone or in combination with other active agents, e.g., fungicides, pesticides, herbicides, antibiotics, antivirals, anesthetics, or plant extracts). Further, the antimicrobial extract may be formulated, withAttorney Docket No.1655.3.WO or without other active agents, as a composition or formulation that includes the antimicrobial extract and an excipient, carrier and / or diluent not present in the original antimicrobial extract. A composition may contain at least 0.001 wt %, 0.05 wt %, 0.1 wt %, 0.5 wt %, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 16 wt%, 17 wt%, 18 wt%, 19 wt%, 20 wt%, 21 wt%, 22 wt%, 23 wt%, 24 wt%, 25 wt%, 26 wt%, 27 wt%, 28 wt%, 29 wt%, 30 wt%, 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35 wt%, 36 wt%, 37 wt%, 38 wt%, 39 wt%, 40 wt%, 41 wt%, 42 wt%, 43 wt%, 44 wt%, 45 wt%, 46 wt%, 47 wt%, 48 wt%, 49 wt%, 50 wt%, 51 wt%, 52 wt%, 53 wt%, 54 wt%, 55 wt%, 56 wt%, 57 wt%, 58 wt%, 59 wt%, 60 wt%, 61 wt%, 62 wt%, 63 wt%, 64 wt%, 65 wt%, 66 wt%, 67 wt%, 68 wt%, 69 wt%, 70 wt%, 71 wt%, 72 wt%, 73 wt%, 74 wt%, 75 wt%, 76 wt%, 77 wt%, 78 wt%, 79 wt%, 80 wt%, 81 wt%, 82 wt%, 83 wt%, 84 wt%, 85 wt%, 86 wt%, 87 wt%, 88 wt%, 89 wt%, 90 wt%, 91 wt%, 92 wt%, 93 wt%, 94 wt%, 95 wt%, 96 wt%, 97 wt%, 98 wt%, 99 wt%, or 100 wt % (or any endpoint or intermediate value within this range) of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, for example, it may contain 0.1-10 wt % of an anhydrous antimicrobial extract and 99.9-90 wt % solvent or other carrier or excipient. Preferably, a composition will contain an effective amount of the antimicrobial extract.

[0057] Compositions containing the antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, may be provided in various forms. Formulations may include other active ingredients and / or non-toxic, inert excipients such as solid, semisolid or liquid diluents, fillers, or adjuvants. Formulations may be in solid, semisolid or viscous, liquid, atomized, aerosol or vaporous forms. They may have an acidic or basic pH and may contain water and / or organic ingredients such as alcohols and other conventional organic excipients, such as those used in pharmaceutical, agricultural, consumer, or cosmetic products. A formulation may be in the form of a solution, tincture, wash (e.g., surgical or dental washes), foam, spray, serum, gel, suppository, suspension, emulsion, cream, lotion, paste, ointment, granule, powder, freeze-dried or desiccated form, troche, capsule, tablet, or pill. An antiseptic or cleaning formulation may contain ingredients conventionally included in such products such as water, organic compounds, chelators, surfactants, oxidants and disinfectants. The particular formulations described in detail below may contain chelators or antioxidants as well as other conventional excipients or carriers. In some aspects, the antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943 may be formulated withAttorney Docket No.1655.3.WO propylene glycol (e.g., 80% by volume extract and 20% by volume propylene glycol), wherein said formulation may be further diluted in water for use in one or more of the methods herein.

[0058] Compositions according to the invention may incorporate one or more chelators that can sequester elements like calcium or iron necessary for bacterial growth. Examples of chelators of iron and calcium include, but are not limited to, diethylene triamine pentaacetic acid (DTPA), ethylene diamine tetraacetic acid (EDTA), nitrilotriacetic acid (NTA), 1,3-propylene diamine tetraacetic acid (PDTA), Ethylene diamine disuccinic acid (EDDS), and ethylene glycol tetraacetic acid (EGTA). Any suitable chelating agent known in the art, which is biologically safe and able to chelate iron, calcium or other metals, is suitable for the invention. Suitable biocompatible chelating agents useful in conjunction with the present invention may include, without limitation, monomeric polyacids such as EDTA, cyclohexanediamine tetraacetic acid (CDTA), hydroxyethylethylenediamine triacetic acid (HEDTA), diethylenetriamine pentaacetic acid (DTPA), dimercaptopropane sulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylene phosphonic acid (ATPA), citric acid, salts thereof, and combinations of any of the foregoing. Other exemplary chelating agents include: phosphates, e.g., pyrophosphates, tripolyphosphates, and hexametaphosphates. Such chelators may be incorporated in amounts sufficient to bind to the divalent cations or metals, such as iron, in a composition. For example, the concentration may be selected to bind to at least 25, 50, 75 or 100 mole % of the calcium, magnesium, or iron in a composition.

[0059] Antioxidants suitable for use in pharmaceutical, cosmetic, food, or agricultural products are known. BHT (butylated hydroxytoluene) and BHA (butylated hydroxyanisole) are two common oil soluble antioxidants. Tocopherols (Vitamin E derivatives, e.g., alpha-tocopherol) and ascorbyl palmitate may also be used. Ascorbates, such as vitamin C, and propyl gallate are examples of water soluble antioxidants. Alpha lipoic acid, acetyl carnitine, Coenzyme Q10 (ubiquinol), selenium, retinoic acid, B vitamins, flavonoids, and various algae and plant extracts may also be used as antioxidants. Such antioxidants may be incorporated in amounts sufficient to quench free radicals in a composition or in a wound to which the composition is applied. For example, the concentration may be selected to bind to at least 25, 50, 75 or 100 mole % of the free radicals in a composition.

[0060] In addition to the active ingredient(s), the formulations described herein, depending on their particular physical and chemical formulation, may contain the customary excipients, forAttorney Docket No.1655.3.WO example animal and vegetable fats, waxes, paraffins, starch, tragant or cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talcum, and zinc oxide, or mixtures of these substances. Aqueous suspensions may contain customary excipients such as liquid diluents, for example water, ethyl alcohol, propylene glycol, suspending agents, for example ethoxylated isostearyl alcohols, polyoxyethylene sorbit and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragant, or mixtures of these substances.

[0061] The kinds of formulations mentioned herein may also contain colorants, preservatives and odor- and flavor-enhancing additives, for example peppermint oil and eucalyptus oil, and, for ingestible products sweeteners and flavorings. Solutions and emulsions according to the invention may contain the customary excipients such as solvents, solubilizers and emulsifiers, for example water, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethyl formamide, oils, in particular cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerin, glycerol formal, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, or mixtures of these substances.

[0062] Some specific kinds of formulations or compositions containing the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, include, but are not limited to, gels, creams, emulsions or suspension concentrates, tablets, aerosols, powders (e.g., wettable powders, dusting powders or soluble powders), pastes, foams, spreading oils or emollients, solutions, sprays, drenches, seed dressings, water-dispersible granules, water soluble granules (slow or fast release), microencapsulated granules or suspensions, or as irrigation components, among others.

[0063] Topical formulations include antimicrobial pharmaceuticals, deodorants, soaps, body washes, shampoos, lubricants (e.g., for catheters or other devices, anatomical sites, or surfaces at risk of microbial contamination), hand or skin sanitizers and disinfectants and personal care products (e.g., a product that is used for personal hygiene). A topical formulation may be provided in a variety of formulations including but not limited to solutions, tinctures, gels, serums, creams, colloids, emulsions, lotions, solid sticks, aerosols or dry powders as described in U.S. Pat. Nos. 4,844,902; 6,818,226; 6,469,015; 7,147,854; 7,192,607; 7,205,003; and 7,252,831, each of which is hereby incorporated by reference for the portions relevant to this paragraph.

[0064] The antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, of the invention may be dissolved in water, another solvent miscible with water, or a mixture thereof forAttorney Docket No.1655.3.WO inclusion in a composition or formulation. The formulation may contain other solutes or liquid components such as salts, chelators (e.g., EDTA, EGTA), antioxidants, or preservatives. In some embodiments, the formulation may further comprise an acid, a base, or buffer for adjusting or stabilizing the pH of a composition so as to maintain or maximize antimicrobial activity of the antimicrobial extract or for suitability for treatment of a particular type of infection. For example, the acid or base is useful for adjusting the pH of a composition to a pH of about 1 to about 14 (e.g., from about 1 to about 2, from about 2 to about 3, from about 3 to about 4, from about 4 to about 5, from about 5 to about 6, from about 6 to about 7, from about 7 to about 8, from about 8 to about 9, from about 9 to about 10, from about 10 to about 11, from about 11 to about 12, from about 12 to about 13, from about 13 to about 14, or any other value or range of values therein). Such pH adjustment may improve the dispersibility of ingredients present in a composition.

[0065] Acids useful in compositions comprising the Vibrio gazogenes extract include inorganic acids such as carbonic acid, sulfuric acid, or hydrochloric acid. Organic acids may alternatively be employed. Suitable organic acids include C1 to C20 organic acids such as formic acid, citric acid, malic acid, adipic acid, tannic acid, lactic acid, ascorbic acid, acetic acid, fumaric acid, and mixtures thereof. In one embodiment, the acid is citric acid. In some embodiments, the compositions do not comprise an acid. These ranges include all intermediate values as well as endpoints.

[0066] Bases useful in compositions comprising the Vibrio gazogenes extract may be organic or inorganic bases. Suitable inorganic bases include alkali metal or alkaline earth metal compounds such as sodium hydroxide, lithium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, magnesium carbonate and calcium carbonate. Other suitable bases include ammonium hydroxide, substituted amine bases and ammonia. In some embodiments, the compositions do not comprise a base.

[0067] In some embodiments, the compositions may comprise one or more salts. Salts useful in compositions comprising the Vibrio gazogenes extract include organic or inorganic salts. Suitable salts include alkali or alkaline earth metal salts such as sodium chloride, sodium nitrate, potassium chloride, calcium chloride, magnesium chloride, ammonium chloride, sodium bromide, potassium bromide, calcium bromide, magnesium bromide, ammonium bromide, sodium iodide, potassium iodide, calcium iodide, magnesium iodide, ammonium iodide, sodium sulfate, potassium sulfate, calcium sulfate, magnesium sulfate, ammonium sulfate. The salt may present in the compositionsAttorney Docket No.1655.3.WO in an amount from 0 wt % to about 30 wt %, e.g., 0 wt % to about 0.5 wt %, about 0.5 wt % to about 1 wt %, about 1 wt % to about 2 wt %, about 2 wt % to about 3 wt %, about 3 wt % to about 4 wt %, about 4 wt % to about 5 wt %, about 5 wt % to about 6 wt %,6 wt % to about 7 wt %, about 7 wt % to about 8 wt %, about 8 wt % to about 9 wt %, about 9 wt % to about 10 wt %, about 10 wt % to about 11 wt %, about 11 wt % to about 12 wt %, about 12 wt % to about 13 wt %, about 13 wt % to about 14 wt %, about 14 wt % to about 15 wt %, about 15 wt % to about 16 wt %, about 16 wt % to about 17 wt %, about 17 wt % to about 18 wt %, about 18 wt % to about 19 wt %, about 19 wt % to about 20 wt %, about 20 wt % to about 21 wt %, about 21 wt % to about 22 wt %, about 22 wt % to about 23 wt %, about 23 wt % to about 24 wt %, about 24 wt % to about 25 wt %, about 25 wt % to about 26 wt %, about 26 wt % to about 27 wt %, about 27 wt % to about 28 wt %, about 28 wt % to about 29 wt %, about 29 wt % to about 30 wt %, or any other value or range of values therein) of the composition. In some embodiments, the salt is present from about 0.01 wt % to about 0.05 wt % of the compositions. In some embodiments, the compositions do not contain a salt.

[0068] A composition may contain an amount of the antimicrobial extract of Vibrio gazogenes that exhibits antimicrobial activity in combination with an amount of solvent sufficient to keep the extract in solution or to dissolve a dry extract. In some embodiments, the amount of solvent (e.g., methanol, ethanol, isopropanol, chloroform, acetonitrile, ether, and / or dimethylsulfoxide) in the compositions comprising the Vibrio gazogenes extract may range from about 10 to about 90 wt % (e.g., about 10 wt % to about 15 wt %, about 15 wt % to about 20 wt %, about 20 wt % to about 25 wt %, about 25 wt % to about 30 wt %, about 30 wt % to about 35 wt %, about 35 wt % to about 40 wt %, about 40 wt % to about 45 wt %, about 45 wt % to about 50 wt %, about 50 wt % to about 55 wt %, about 55 wt % to about 60 wt %, about 60 wt % to about 65 wt %, about 65 wt % to about 70 wt %, about 70 wt % to about 75 wt %, about 75 wt % to about 80 wt %), about 80 wt % to about 85 wt %, about 85 wt % to about 90 wt %, or any other value or range of values therein). In certain embodiments, compositions comprising the Vibrio gazogenes extract comprise from about 80 wt % to about 90 wt % water or about 90 wt. % to about <100 wt. % water.

[0069] The compositions of the invention may further contain one or more other additives. Suitable additives include, but are not limited to, detergents, as surface tension modifiers, flocculants, dispersants, rheology modifiers, emulsifiers, surfactants, chelators, and solvents. Illustrative additives are polysorbates, oils (e.g., canola oil, vegetable oils, etc.). In someAttorney Docket No.1655.3.WO embodiments, the additive(s) may be present in the compositions in amounts ranging from 0 to about 30 wt % (e.g., 0 to about 0.5 wt %, about 0.5 wt % to about 1 wt %, about 1 wt % to about 2 wt %, about 2 wt % to about 3 wt %, about 3 wt % to about 4 wt %, about 4 wt % to about 5 wt %, about 5 wt % to about 6 wt %, about 6 wt % to about 7 wt %, about 7 wt % to about 8 wt %, about 8 wt % to about 9 wt %, about 9 wt % to about 10 wt %, about 10 wt % to about 11 wt %, about 11 wt % to about 12 wt %, about 12 wt % to about 13 wt %, about 13 wt % to about 14 wt %, about 14 wt % to about 15 wt %, about 15 wt % to about 16 wt %, about 16 wt % to about 17 wt %, about 17 wt % to about 18 wt %, about 18 wt % to about 19 wt %, about 19 wt % to about 20 wt %, about 20 wt % to about 21 wt %, about 21 wt % to about 22 wt %, about 22 wt % to about 23 wt %, about 23 wt % to about 24 wt %, about 24 wt % to about 25 wt %, about 25 wt % to about 26 wt %, about 26 wt % to about 27 wt %, about 27 wt % to about 28 wt %, about 28 wt % to about 29 wt %, about 29 wt % to about 30 wt %, or any other value or range of values therein) of the composition.

[0070] In some embodiments, compositions of the invention may contain a surfactant. Surfactants are compounds that lower the surface tension of a liquid, the interfacial tension between two liquids, or that between a liquid and a solid. Surfactants may act as detergents, wetting agents, emulsifiers, foaming agents, and dispersants. Surfactants that may be present in the compositions of the invention include anionic surfactants, amphoteric surfactants, cationic surfactants, zwitterionic surfactants, non-ionic surfactants, and combinations thereof. Surfactants suitable for use in the present invention may include polysorbates such as polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80. An amount of surfactant that is sufficient to emulsify other ingredients in a composition or that is sufficient to lower surface tension of a composition so as to maximize contact between a surface and the antimicrobial extract or maximize antimicrobial activity may be selected.

[0071] Gels provided herein include semi-solid suspensions that contain the antimicrobial extract. The gels may be single- or two-phase systems. The gels may be oil or liquid based. Single- phase gels may contain small organic macromolecules distributed substantially uniformly throughout a liquid, such that there is no boundary between the macromolecules and liquid. The liquid may be aqueous, but also contain an alcohol, and, optionally, an oil. Single-phase gels may be made from synthetic macromolecules or from natural gums. Two-phase gels may include a network of small, discrete particles. In one embodiment, two-phase gels are thixotropic. In oneAttorney Docket No.1655.3.WO embodiment, the organic macromolecules include crosslinked acrylic acid polymers such as the “carbomer” family of polymers (i.e., carboxypolyalkylenes). The organic macromolecules may also be hydrophilic polymers such as polyethylene oxides, polyoxyethylene-polyoxypropylene copolymers and polyvinyl alcohol; cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methyl cellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin. In some embodiments, the organic macromolecules having a stabilizing action include long-chain linear high molecular weight polysaccharides with a molecular weight of more than one million. In this embodiment, 0.1 wt% to 1.5 wt % of such stabilizers may be included. In some embodiments, a uniform gel may be prepared by adding dispersing agents such as alcohol or glycerin. In some embodiments, the organic macromolecules may be dispersed by trituration, mechanical mixing or stirring, or combinations thereof. In some embodiments, the liquid maybe either water or a water-miscible solvent.

[0072] Creams provided herein include liquids or semi-solid emulsions with a viscous consistency. Creams maybe either oil-in-water or water-in-oil based formulations. Cream bases maybe water soluble. Cream bases may contain the following components: (1) an oil phase, (2) an aqueous phase, and (3) an emulsifier. The oil phase may comprise petroleum jelly and a fatty alcohol, such as cetyl or stearyl alcohol. The aqueous phase may contain a humectant. The emulsifier maybe a nonionic, anionic, cationic or amphoteric surfactant. In one embodiment, the oil phase includes, but is not limited to, cetyl alcohol, stearyl alcohol, stearic acid, liquid paraffin, and dimethicone. In some embodiments, the water phase ingredient includes, but is not limited to, glycerol and ethyl paraben as well as the antimicrobial extract. In some embodiments, the emulsifying agent includes, but is not limited to, fatty alcohol polyoxyethylene ether (PEREGAL™ A-20), polyoxylstearate (SG-6), or combinations thereof.

[0073] Lotions provided herein include liquids or semi-liquid formulations that are generally lower in viscosity than a cream or gel. The lotions maybe an oil-in-water formulation stabilized by a surface-active agent and are usually suitable for application to unbroken skin. In one embodiment, the lotions contain suspending agents to produce better dispersions and compounds useful for localizing and holding the active agent such as components of the antimicrobial extract in contact with the skin, including methylcellulose, sodium carboxymethyl-cellulose, and similar compounds.Attorney Docket No.1655.3.WO

[0074] Ointments provided herein include semi-solid preparations that have petroleum jelly or their derivatives as a base. Petroleum jelly is a semi-solid mixture of hydrocarbons. As described in Remington: The Science and Practice of Pharmacy, 19th Ed. (Easton, Pa.: Mack Publishing Co., 1995), ointment bases may be grouped in four classes: oleaginous bases; emulsifiable bases; emulsion bases; and water-soluble bases. Oleaginous ointment bases include vegetable oils, fats obtained from animals, and semisolid hydrocarbons obtained from petroleum. Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and include, for example, hydroxystearin sulfate, anhydrous lanolin, and hydrophilic petroleum jelly. Emulsion ointment bases are either water-in-oil or oil-in-water emulsions, and include, for example, cetyl alcohol, glyceryl monostearate, lanolin, and stearic acid. An ointment may contain solid or encapsulated particles of a dry antimicrobial extract of Vibrio gazogenes or may contain an emulsified or suspended antimicrobial extract of Vibrio gazogenes.

[0075] Pastes included herein contain, in addition to an ointment or cream base, high amounts of pulverulent constituents, such as zinc oxide, talc, starch or titanium dioxide. In one embodiment, the paste is selected from the group comprising fatty pastes or single-phase aqueous gels. The fatty paste includes petroleum jelly, hydrophilic petroleum jelly, or other similar compounds. The single-phase aqueous gel may incorporate carboxymethylcellulose or similar compounds. A paste may contain solid or encapsulated particles of a dry antimicrobial extract of Vibrio gazogenes or may contain an emulsified or suspended antimicrobial extract of Vibrio gazogenes.

[0076] Aerosol as provided herein includes products packaged under pressure and contain ingredients that are released upon activation of an appropriate valve system. Aerosols include all self-contained pressurized products, such as fine mists of spray or foam, that are emitted from a pressurized container containing a propellant, foams, or semisolid liquids. They may also be emitted by an unpressurized atomizer that is pressurized by a hand-operated pump rather than by stored propellant. In one embodiment, the aerosol comprises a container, a propellant, a concentrate containing an active ingredient, a valve (which may be a metered valve), and an actuator. The nature of these components determines characteristics such as delivery rate, foam density, and fluid viscosity. In some embodiments, the aerosol is a two-phase formulation comprising a gas and liquid. In some embodiments, the aerosol is a three-phase formulation comprising a gas, liquid, and suspension or emulsion of active ingredients. In this formulation, suitable excipients, such as wetting agents and / or solid carriers such as talc or colloidal silicas areAttorney Docket No.1655.3.WO included. In some embodiments, the propellant is liquefied or vaporized. In some embodiments, a solvent maybe the propellant or a mixture of the propellant and co-solvents such as alcohol and polyethylene glycols. In some embodiments, the propellant is selected from the group comprising a spray, foam, or quick-breaking foam. In some embodiments, spray formulations are aqueous solutions in a container having a spray means, such as an atomizer or nebulizer. A spray may contain an aerosol of solid or encapsulated particles of a dry antimicrobial extract of Vibrio gazogenes or may contain an emulsified or suspended antimicrobial extract of Vibrio gazogenes in an atomized or aerosol liquid form.

[0077] A composition according to the invention may also be atomized or aerosolized for dispersement within a space, such as in a hospital or a room in which a patient recovering from a microbial infection, e.g., pneumonia, is housed. Methods for dispersement of such compositions are described by and incorporated by reference to Schur, U.S. Pat. No.7,638,114.

[0078] In some embodiments, the antimicrobial extract is delivered to the body while in a foam state, such as stable foam, for example, that is produced with or without a propellant. For example, the antimicrobial extract may be contained in a shaving foam and used for preventing bacterial infection of nicks, cuts or abrasions associated with shaving. In some versions, a foam is dispensed from a dispenser such as a propellant-free dispenser with pumping action to create the foam from a composition in a foamable carrier, and then applied to a wipe or other substrate, or applied to the hand of the user or otherwise delivered to the skin. Propellant-driving foam generators may also be used to deliver the composition in the form of a foam. Active ingredients in a foam may be dispensed for subsequent placement on a dry wipe, a pre-moistened wipe, or other soft, flexible applicator (e.g., an object about 3-fingers wide or 4 to 10 cm wide) or other object to be used for application of the foam-based composition to the skin. The foam maybe a non-propellant foam. A foam with a suitable stiffness of yield stress may be applied to the skin in any manner for sustained adherence and contact with the body. Examples of foam-based systems are described in U.S. Pat. No.6,818,204. At least one foam stabilizing agent may be present in some foamable embodiments. Suitable foam stabilizing agents may include, without limitation, natural or synthetic gums such as xanthan gum, polyalkylene glycols such as polyethylene glycol, alkylene polyols such as glycerine and propylene glycol and combinations thereof. Typically, the foam stabilizers may be present in an amount from about 0.10% to about 5%, or from about 2% to about 4%.Attorney Docket No.1655.3.WO

[0079] The topical formulations provided herein may include additional ingredients to affect the physical or functional characteristics of the formulations. Stabilizers, preservatives, humectants, regreasing agents, solvents or auxiliaries may be included to improve efficacy and dermal penetration. Dermal penetration-enhancing compounds provided have low toxicity to the skin and may promote percutaneous and oral mucosal absorption. In one embodiment, dermal penetration- enhancing compounds include propylene glycol, polyethylene glycol, dimethyl sulphoxide, decylmethylsulphoxide, azones, N-methylpyrrolidone, diethyltoluamide, ethanol, isopropyl myristate, isopropyl palmitate, oleic acid and its esters, medium-chain triglycerides, dimethyl isosorbitol, 2-octyldodecanol, branched fatty acids, benzyl alcohol, urea, salicylates and surfactants. Viscosity enhancers or thickeners may be included. Such enhancers may prevent the formulation from spreading beyond the site of application. In one embodiment, Balsam Fir is a pharmaceutically acceptable viscosity enhancer. Another benefit of increasing the viscosity of the formulation is provided below in the section discussing thixotropic agents. Thickeners include suitable polymers such as carbomer, hydroxypropyl methylcellulose, hydroxyethylcellulose, PVM / MA decadiene cross-polymer and acrylates. Two or more thickeners may be added.

[0080] Spreading oils or emollients may be included in topical formulations. One benefit for including such oils is for better distribution on surfaces. Spreading oils are understood as those oily liquids which are distributed particularly easily on the skin. They are known as such in cosmetics. The following compounds are suitable spreading agents: silicone oil, fatty acid esters, such as ethyl stearate, di-n-butyl adipate, hexyl laurate and dipropylene glycol pelargonate, esters of a branched fatty acid of medium chain length with saturated C16-C18 fatty alcohols, isopropyl myristate, isopropyl palmitate, caprylic / capric acid esters of saturated fatty alcohols of C12-C18chain length, isopropyl stearate, oleyl oleate, decyl oleate, ethyl oleate, ethyl lactate, waxy fatty acid esters, such as synthetic duck uropygial gland fat, dibutyl phthalate, diisopropyl adipate, ester mixtures related to the latter and the like. Other elements that may be included are emollients, such as diisopropyl adipate / isohexadecane dimethicone, occlusive agents, such as cyclomethicone, trimethylsiloxysilicate, glycereth-26 or polyquaternium-7, emulsifiers, such as cetyl alcohol, stearyl, stearic acid, glyceryl stearate, propylene glycol isostearoyl-sodium isostearoyl, a lactylate, polyoxyethylene (100) stearate, skin conditioners, moisturizers, humectants, such as propylene glycol or glycerin, preservatives, such as phenoxyethanol and parabens, pH adjusting agents,Attorney Docket No.1655.3.WO surfactants, chelators, such as disodium EDTA or sodium citrate, tackifying agents, fragrances and other compounds.

[0081] In some embodiments, the antimicrobial extract of Vibrio gazogenes may be formulated as a tablet, capsule or pill that contains the antimicrobial extract of Vibrio gazogenes. These may contain the customary excipients, such as fillers and extenders, for example starches, lactose, sucrose, glucose, mannitol, and silicic acid; binders, for example carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone; humectants, for example glycerin; disintegrating agents, for example agar-agar, calcium carbonate and sodium carbonate; dissolution retardants, for example paraffin; resorption accelerating agents, for example quaternary ammonium compounds; wetting agents, for example cetyl alcohol, glycerol monostearate; adsorption agents, for example kaolin and bentonite; and lubricants, for example talcum, calcium stearate and magnesium stearate, and solid polyethylene glycols or mixtures of the substances mentioned above. In some embodiments, the active ingredient(s) can be in a microencapsulated form in the tablet or capsule, which can optionally be formulated to release the active antimicrobial extract component at a particular location within the gastrointestinal tract, e.g, to transit the stomach and release the active component in the small or large intestine.

[0082] A composition according to the invention may be formulated in the form of an antimicrobial powder which contains a dry or encapsulated antimicrobial extract of Vibrio gazogenes and the customary excipients, for example lactose, talcum, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these substances. Such powders may be formulated for topical application or for inhalation.

[0083] The compositions of the invention can be formulated as deodorants or personal care products that prevent the formation of body odors such as those produced by the growth of Gram- positive bacteria in or on the skin. Such deodorant will contain an effective amount of the antimicrobial extract of Vibrio gazogenes and suitable excipients or carriers to facilitate its application to the body. The antimicrobial extract of Vibrio gazogenes may be incorporated into conventional body washes, lotions, lubricants, personal care composition, antiperspirants, or deodorants. Such products are well known in the art and commercially available and are also described by Broad, U.S. Pat. No. 4,252,789, which is incorporated by reference, especially for their descriptions of conventional deodorant ingredients, formulations, and modes of use. These products can be applied to the axilla, inguinal region, feet or other odor-producing body part toAttorney Docket No.1655.3.WO prevent growth of odor-causing microorganisms. In some embodiments, the antimicrobial extract of the invention may be incorporated into a deodorizer, cleaner, or disinfectant such as a liquid sanitizer or disinfectant, a spray or wipe for cleaning surfaces exposed to bacterial contaminants.

[0084] A composition containing the antimicrobial extract of Vibrio gazogenes may be formulated for use as a cleaning or disinfecting agent, such as a hard surface cleaning product. It may further contain cleaning agents, such as chelators or surfactants, which do not interfere with the antimicrobial activity of the extract. The formulation of such a cleaning or disinfecting solution and inclusion of general cleaning agents can easily be done by a skilled artisan and the stability and effectiveness of the solution can be easily tested by the skilled artisan. The term “hard surface cleaning composition” refers to a composition that is used to clean and / or sanitize a hard or solid inanimate surface. In one embodiment, the invention provides a composition that prevents microbial growth on hard surfaces including, but not limited to, surgical instruments, storage tanks, pipelines, trays, containers, walls, floors, countertops, locker room floors, benches, lockers, showers, bathrooms, toilets, water filtration units, and the like.

[0085] In other aspects, the antimicrobial extract of Vibrio gazogenes extract of the invention provided in the form of an ingestible food or beverage or in an additive or protectant in aquaculture. In some embodiments, the ingestible food or beverage may be a human food or animal feed that contains the antimicrobial extract of Vibrio gazogenes. The antimicrobial extract of Vibrio gazogenes may be incorporated into animal feeds for mammals (cattle, sheep, goats, etc.), birds (e.g., chickens, turkeys, quail, ducks, geese, hawks, falcons, etc.), fish (e.g., tilapia, carp, catfish, salmon, trout, aqua cultured fish, etc.), and crustaceans (e.g., shrimp, lobsters, etc.), mollusks (e.g., abalone, oysters, clams, mussels, etc.). In some embodiments, the antimicrobial extract of the invention may be incorporated into a liquid medium in which an animal is grown, e.g., into a medium for aquaculture. In some embodiments, the extract may be encapsulated in a form that permits uptake by an animal, for example, in an encapsulated particulate form that can be ingested by fish.

[0086] In some aspects, the Vibrio gazogenes extract of the invention may be applied to control the growth of microbes in or on plants, such those causing root rot, leaf spot, ear rots, wilt, or powdery mildew. It may be sprayed or otherwise applied to any plant or part thereof, such as, for example, the roots, foliage, flowers, fruit or seeds. In some embodiments, the extract may be added to culture medium used for hydroponic cultivation of plants.Attorney Docket No.1655.3.WO

[0087] In an additional aspect, the present invention provides a commodity, industrial, or commercial product comprising the antimicrobial extract of the invention. In some embodiments, the antimicrobial extract of V. gazogenes, ATCC Accession No. 43943, as described herein is substantially devoid of membrane components and comprises compounds soluble in (a) methanoland (b) acetone, ethanol, chloroform, acetonitrile, or dimethylsulfoxide. In some embodiments, the antimicrobial extract comprises prodigiosin, 6-hydroxycaproic acid, acetophenone, 2'-deoxyadenosine, baquiloprim and peptides. Examples of products that may include the antimicrobial extract include, but are not limited to, (a) a pharmaceutical composition suitable for administration orally, dermally, parenterally, nasally, ophthalmically, otically, sublingually, rectally and / or vaginally; (b) a food product including a fish, crustacean, fish substitute, crustacean substitute, meat, meat substitute, poultry product, vegetable, green, sauce, emulsion, beverage, juice, wine, beer, dairy product, egg-based product, jam, jelly, grain-based product, baked good or confectionary product; (c) a cosmetic or personal care product including a cream, gel, powder, lotion, sunscreen, lipstick, body wash, herbal extract, or formulation that supports the growth of bacteria; (d) an agricultural product including a foliar spray, root drench, seed coating, or animal feed; or (e) a surface cleaning product including a cleaning product for counter tops, doors, windows, handles, surgical equipment, or medical tools.

[0088] The antimicrobial extract of the Vibrio gazogenes strain available under ATCC Accession No.43943 and biologically pure isolates thereof have activity against a broad range of microbes. In one aspect, the antimicrobial extract of Vibrio gazogenes, Accession No. 43943, has antimicrobial activity against microbial pathogens. In another aspect, the antimicrobial extract of Vibrio gazogenes, Accession No.43943, has antimicrobial activity against fungi and / or bacteria. In a further aspect, the antimicrobial extract of Vibrio gazogenes, Accession No. 43943, has antimicrobial activity against pathogenic fungi and / or pathogenic bacteria. In some aspects, the IC50 value of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, toward a microbial pathogen is in the range of about 50 ppm to about 200 ppm (e.g., 50 ppm, 51 ppm, 52 ppm, 53 ppm, 54 ppm, 55 ppm, 56 ppm, 57 ppm, 58 ppm, 59 ppm, 60 ppm, 61 ppm, 62 ppm, 63 ppm, 64 ppm, 65 ppm, 66 ppm, 67 ppm, 68 ppm, 69 ppm, 70 ppm, 71 ppm, 72 ppm, 73 ppm, 74 ppm, 75 ppm, 76 ppm, 77 ppm, 78 ppm, 79 ppm, 80 ppm, 81 ppm, 82 ppm, 83 ppm, 84 ppm, 85 ppm, 86 ppm, 87 ppm, 88 ppm, 89 ppm, 90 ppm, 91 ppm, 92 ppm, 93 ppm, 94 ppm, 95 ppm, 96 ppm, 97 ppm, 98 ppm, 99 ppm, 100 ppm, 101 ppm, 102 ppm, 103 ppm, 104 ppm, 105 ppm, 110Attorney Docket No.1655.3.WO ppm, 111 ppm, 12 ppm, 113 ppm, 114 ppm, 115 ppm, 116 ppm, 117 ppm, 118 ppm, 119 ppm, 120 ppm, 121 ppm, 122 ppm, 123 ppm, 124 ppm, 125 ppm, 126 ppm, 127 ppm, 128 ppm, 129 ppm, 130 ppm, 131 ppm, 132 ppm, 133 ppm, 134 ppm, 135 ppm, 140 ppm, 141 ppm, 142 ppm, 143 ppm, 144 ppm, 145 ppm, 146 ppm, 147 ppm, 148 ppm, 149 ppm, 150 ppm, 151 ppm, 152 ppm, 153 ppm, 154 ppm, 155 ppm, 156 ppm, 157 ppm,158 ppm, 159 ppm, 160 ppm, 161 ppm, 162 ppm, 163 ppm, 164 ppm, 165 ppm, 170 ppm, 171 ppm, 172 ppm, 173 ppm, 174 ppm, 175 ppm, 180 ppm, 185 ppm, 186 ppm, 187 ppm, 188 ppm, 189 ppm, 190 ppm, 191 ppm, 192 ppm, 193 ppm, 194 ppm, 195 ppm, 196 ppm, 197 ppm, 198 ppm, 199 ppm, or 200 ppm, or any range or value therein).

[0089] The present invention further provides a method for preventing or reducing the growth of a microbial pathogen comprising contacting a microbial pathogen with an effective amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943 as described herein. In some embodiments, the antimicrobial extract of V. gazogenes, ATCC Accession No. 43943, as described herein is substantially devoid of membrane components and comprises compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide. In some embodiments, the antimicrobial extract comprises prodigiosin, 6- hydroxycaproic acid, acetophenone, 2'-deoxyadenosine, baquiloprim and peptides. In some aspects of the disclosure, the microbial pathogen is a pathogenic fungus such as pathogenic Aspergillus spp. (e.g., A. flavus, A. parasiticus, A. fumigatus), Rhizoctonia spp. (e.g., R. solani), Candida spp. (e.g, C. albicans), Podosphaera spp. (e.g., P. aphanis), Penicillium spp. (e.g., P. expansum), Botrytis spp. (e.g., B. cinerea), Verticillium spp. (e.g., V. dahlia), Fusarium spp. (e.g., F. oxysporum), Phoma spp. (e.g., P. betae), Cersospora spp., Magnaporthe spp, or Phyllachora spp. In other aspects of the disclosure, the microbial pathogen is a pathogenic bacterium such as pathogenic Leuconostoc spp. (e.g., L. mesenteroides), Escherichia spp. (e.g., E. coli), Staphylococcus spp. (e.g., S. aureus), Pseudomonas spp. (e.g., P. aeruginosa), Porphyromonas spp. (e.g., P. gingivalis), Enterococcus spp. (e.g., E. faecium), Salmonella spp., or Listeria spp. (e.g., L. monocytogenes). Growth of a microbial pathogen may be assessed by conventional assays including, but not limited to, liquid culture-based assays and plate-based assays as described herein.

[0090] The present invention also provides a method for preventing or reducing the likelihood of contamination of a food product by a microbial pathogen comprising applying an effectiveAttorney Docket No.1655.3.WO amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, as described herein to the food product. In some embodiments, the antimicrobial extract of V. gazogenes, ATCC Accession No.43943, as described herein is substantially devoid of membrane components and comprises compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide. In some embodiments, the antimicrobial extract comprises prodigiosin, 6-hydroxycaproic acid, acetophenone, 2'- deoxyadenosine, baquiloprim and peptides. In some aspects, the food product may be a beverage, vegetable, fruit, and / or processed food. The antimicrobial extract may be applied to the surface of the food product (e.g., the surface of fruits or vegetables), may be added directly to the food product during processing, or applied onto the packaging of the food product.

[0091] A method for inhibiting the growth of a microbial pathogen on an inanimate surface or environmental sample is also provided, wherein the method includes the step of contacting the inanimate surface or environmental sample with an effective amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, as described herein, thereby inhibiting the growth of a microbial pathogen. In some embodiments, the antimicrobial extract of V. gazogenes, ATCC Accession No.43943, as described herein, is substantially devoid of membrane components and comprises compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide. In some embodiments, the antimicrobial extract comprises prodigiosin, 6-hydroxycaproic acid, acetophenone, 2'-deoxyadenosine, baquiloprim and peptides. Examples of inanimate surfaces include, but are not limited to, countertops, desks, carts, trays, tables, chairs, medical equipment, and building materials (e.g., paints and wood). Examples of environmental samples include, e.g., soil samples and water samples. In some aspects, growth of the microbial pathogen on the inanimate surface or environmental sample may be inhibited by about 50% to about 90% (e.g., about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90%, or any range or value therein) as compared to a control.

[0092] The present invention further provides a method for preventing, treating, or reducing the likelihood of infection by a microbial pathogen (e.g, a pathogenic fungus or bacterium) comprising administering to a subject in need thereof an effective amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No. 43943, as described herein. In some embodiments, theAttorney Docket No.1655.3.WO antimicrobial extract of V. gazogenes, ATCC Accession No. 43943, as described herein is substantially devoid of membrane components and comprises compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide. In some embodiments, the antimicrobial extract comprises prodigiosin, 6-hydroxycaproic acid, acetophenone, 2'-deoxyadenosine, baquiloprim and peptides. Subjects benefiting from treatment with the antimicrobial extract include subjects with a burn or wound (e.g., a gunshot wound, a puncture wound, a laceration wound, an abrasion, a cut, a penetration wound, a surgical wound, or any other wound) as well as subjects that have inhaled or been suspected of inhaling a microbial pathogen, ingested contaminated food or water; or contacted a contaminated surface. In some aspects, the antimicrobial extract is used in the prevention, treatment, or reduction in the likelihood of an infection by a pathogenic bacterium such as E. coli, Salmonella, Listeria (e.g., L. monocytogenes), S. aureus, P. areuginosa, and / or P. gingivalis. In other aspects, the antimicrobial extract is used in the prevention, treatment, or reduction in the likelihood of aspergillosis or candidiasis caused by an infection with Aspergillus (e.g., A. fumigatus) and / or Candida (e.g, C. albicans), respectively.

[0093] In another aspect, the invention provides a method for treating, preventing or reducing the likelihood of infection of a plant or part thereof by a microbial pathogen comprising contacting the plant or part thereof with an effective amount of the antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, as described herein. In some embodiments, the antimicrobial extract of V. gazogenes, ATCC Accession No. 43943, as described herein, is substantially devoid of membrane components and comprises compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide. In some embodiments, the antimicrobial extract comprises prodigiosin, 6-hydroxycaproic acid, acetophenone, 2'- deoxyadenosine, baquiloprim and peptides. All plants and plant parts may be treated in accordance with the invention. In the present context, plants are understood as meaning all plants and plant populations, such as desired and undesired wild plants or crop plants (including naturally occurring crop plants), and may include a plant and / or plant part of any plant species / variety / cultivar. A plant may include, but is not limited to, an angiosperm, a gymnosperm, a monocot, a dicot, a C3 plant, a C4 plant, a CAM plant, a bryophyte, a fern and / or fern ally, microalgae, and / or macroalgae. Crop plants may be plants which may be obtained by traditional breeding and optimization methods or by biotechnological and recombinant methods, or combinations of these methods. Plant parts areAttorney Docket No.1655.3.WO understood as meaning all aerial and subterranean parts and organs of the plants, including but not limited to a shoot, leaf, flower and root, examples which may be mentioned being leaves, needles, stalks, stems, flowers, fruiting bodies, fruits and seeds, and also roots, tubers and rhizomes. The plant parts also include any crop or plant material (e.g., tissue culture material including cells, protoplasts, or callus tissue) and any vegetative and generative propagation material, for example cuttings, tubers, rhizomes, slips and seeds. Non-limiting examples of plants that may be contacted with the Vibrio gazogenes extract as described herein, and compositions and formulations thereof may include, but are not limited to, turf grasses (e.g., bluegrass, bentgrass, ryegrass, fescue), feather reed grass, tufted hair grass, miscanthus, arundo, switchgrass, vegetable crops, including artichokes, kohlrabi, arugula, leeks, asparagus, lettuce (e.g., head, leaf, romaine), malanga, melons (e.g., muskmelon, watermelon, crenshaw, honeydew, cantaloupe), cole crops (e.g., brussels sprouts, cabbage, cauliflower, broccoli, collards, kale, chinese cabbage, bok choy), cardoni, carrots, napa, okra, onions, celery, parsley, chick peas, parsnips, chicory, peppers, potatoes, cucurbits (e.g., marrow, cucumber, zucchini, squash, pumpkin, honeydew melon, watermelon, cantaloupe), radishes, dry bulb onions, rutabaga, eggplant, salsify, escarole, shallots, endive, garlic, spinach, green onions, squash, greens, beet (sugar beet and fodder beet), sweet potatoes, chard, horseradish, tomatoes, turnips, and spices; a fruit crop such as apples, apricots, cherries, nectarines, peaches, pears, plums, prunes, cherry, quince, fig, nuts (e.g., chestnuts, pecans, pistachios, hazelnuts, pistachios, peanuts, walnuts, macadamia nuts, almonds, and the like), citrus (e.g., clementine, kumquat, orange, grapefruit, tangerine, mandarin, lemon, lime, and the like), blueberries, black raspberries, boysenberries, cranberries, currants, gooseberries, loganberries, raspberries, strawberries, blackberries, grapes (wine and table), avocados, bananas, kiwi, persimmons, pomegranate, pineapple, tropical fruits, pomes, melon, mango, papaya, and lychee, a field crop plant such as clover, alfalfa, timothy, evening primrose, meadow foam, corn / maize (field, sweet, popcorn), hops, jojoba, buckwheat, safflower, quinoa, wheat, rice, barley, rye, millet, sorghum, oats, triticale, sorghum, tobacco, kapok, a leguminous plant (beans (e.g., green and dried), lentils, peas, soybeans), an oil plant (rape, canola, mustard, poppy, olive, sunflower, coconut, castor oil plant, cocoa bean, groundnut, oil palm), duckweed, Arabidopsis, a fiber plant (cotton, flax, hemp, jute), Cannabis (e.g., Cannabis sativa, Cannabis indica, and Cannabis ruderalis), lauraceae (cinnamon, camphor), or a plant such as coffee, sugar cane, tea, and natural rubber plants; and / or a bedding plant such as a flowering plant, a cactus, a succulent and / or anAttorney Docket No.1655.3.WO ornamental plant (e.g., roses, tulips, violets), as well as trees such as forest trees (broad-leaved trees and evergreens, such as conifers; e.g., elm, ash, oak, maple, fir, spruce, cedar, pine, birch, cypress, eucalyptus, willow), as well as shrubs and other nursery stock.

[0094] In some aspects, application of the antimicrobial extract to a plant or part thereof may be used in the control of plant diseases such as root rot, leaf spot, ear rots, fusarium wilt, powdery mildew infestations in plants including, but not limited to, corn, sugar beet, sweet potato, potato, tomato, berries, banana, apples, squash, coffee, spinach, and broccoli. Accordingly, the antimicrobial extract may be useful against plant pathogenic bacteria such as P. aeruginosa and plant pathogenic fungi such as Aspergillus spp. (e.g., A. flavus, A. parasiticus, A. fumigatus), Rhizoctonia spp. (e.g., R. solani), Podosphaera spp. (e.g., P. aphanis), Penicillium spp. (e.g., P. expansum), Botrytis spp. (e.g., B. cinerea), Verticillium spp. (e.g., V. dahlia), Fusarium spp. (e.g., F. oxysporum), Phoma spp. (e.g., P. betae), Cersospora spp., Magnaporthe spp, or Phyllachora spp.

[0095] Depending on the intended recipient (e.g., a subject, plant, food product, environmental sample, or inanimate surface), application of the antimicrobial extract may be carried out directly or by acting on the environment or storage space using customary treatment methods, for example by wound dressing, dipping, spraying, atomizing, misting, evaporating, dusting, fogging, scattering, foaming, painting on, spreading, injecting, drenching, trickle irrigation and, in the case of plant propagation material, in particular in the case of seed, furthermore by the dry seed treatment method, the wet seed treatment method, the slurry treatment method, by encrusting, by coating with one or more coats and the like. It is furthermore possible to apply the antimicrobial extract by the ultra-low volume method or to inject the active substance preparation or the active substance itself into the soil. By way of illustration, the antimicrobial extract may be initially suspended in a small volume of, e.g., ethanol, methanol, or DMSO, just enough to cover the extract, thoroughly mixed, and then applied at a concentration in the range of 100 ng to 1 mg of extract per square inch of area to be treated (e.g., an infected surface or substrate).

[0096] The invention will now be described with reference to the following examples. It should be appreciated that these examples are not intended to limit the scope of the claims to the invention, but are rather intended to be exemplary of certain embodiments. Any variations in the exemplified methods that occur to the skilled artisan are intended to fall within the scope of the invention.Attorney Docket No.1655.3.WO EXAMPLES Example 1. Screening of Vibrio gazogenes Strains for Antimicrobial Activity

[0097] Vibrio gazogenes, Accession Nos. 43939 through 43944, were obtained from ATCC. Vibrio gazogenes, Accession No.43943 is described by Farmer et al. ((1998) J. Clin. Microbiol. 26(10):1993-2000). Marine culture medium was used for culturing these strains. The contents of marine medium per liter are: Peptone (5.0 g), Yeast Extract (1.0 g), Ferric Citrate (0.1 g), Sodium Chloride (19.45 g), Magnesium Chloride (5.9 g), Magnesium Sulfate (3.24 g), Calcium Chloride (1.8 g), Potassium Chloride (0.55 g), Sodium Bicarbonate (0.16 g), Potassium Bromide (0.08 g), Strontium Chloride (34.0 mg), Boric Acid (22.0 mg), Sodium Silicate (4.0 mg), Sodium Fluoride (2.4 mg), Ammonium Nitrate (1.6 mg), and Disodium Phosphate (8.0 mg). The cells were incubated for 18-24 hours at 26°C, centrifuged, and the resulting pellets were extracted with 1 mL of 99% methanol by shaking for 30 minutes at 25°C. Acetone (5 mL) was subsequently added and the resulting mixture was shaken for another 30 minutes. The solvent mixture containing the cellular extract was dried to obtain cellular extracts that were screened for antimicrobial activity against Aspergillus flavus. Based upon colony diameter measurements, it was observed that compared to ATCC Accession No.43942, Accession No.43943 inhibited the growth of A. flavus by 50-60% (FIG.1).

[0098] LC-MS / MS analysis of the extracts from Vibrio gazogenes, Accession Nos. 43939 through 43944, indicated that while these natural isolates were grown under identical growth conditions, each produced a very different chromatographic profile of compounds (FIG.2). For example, the chromatogram profile of extracts obtained from ATCC 43942 and that obtained from ATCC 43943 were measurably different.

[0099] Vibrio gazogenes strain ATCC 43942 is a strain known to inhibit mycotoxin production. See US 2022 / 0249576 A1. However, antimicrobial activity has not been previously demonstrated. Therefore, to determine whether the lack of antimicrobial activity is due to an absence of inhibitory compounds or just an effect of dose, a dose-response experiment was conducted wherein Aspergillus flavus growth was quantified using dry weight measurements of mycelia harvested 2 days post inoculation. The results (FIG. 3) indicate that when extracts from ATCC 43942 were applied at 3-fold higher dose relative to the dosage applied for ATCC 43943, the fungal growth significantly increased and was not inhibited. Therefore, ATCC 43942 does not appear to exhibit antimicrobial activity.Attorney Docket No.1655.3.WO Example 2. Preparation of Antimicrobial Extract

[0100] Vibrio gazogenes, Accession No.43943, was single colony isolated by streaking a frozen stock culture onto a marine agar plate, incubating the plate at 26°C until single colonies were visible, and selecting a single red colony from the plate. The single colony was needle inoculated into 5 mL of marine medium and incubated at 26°C for 18-24 hours. One mL of the overnight culture was then transferred into 100 mL of fresh marine medium and incubated an additional 18- 24 hours at 26°C or until an OD600 of 1.2 was reached. Prior to extraction of the entire culture, 1.5 mL of the culture was removed, the sample was centrifuged to obtain a cellular pellet, the cellular pellet was resuspended in 500 mL methanol with vortexing, and the resulting extract was analyzed by spectrophotometry to confirm that the cells produced compounds having peaks at 465 nm, 505 nm, and 535 nm (FIG.4).

[0101] After growing the bacteria for 18-24 hours, the bacterial cells were harvested by centrifugation. The bacterial pellet was then resuspended in 1 mL of 100% methanol with shaking to disrupt the bacterial membrane and kill the bacteria. After a 30 minute incubation in methanol, 5 mL of acetone was added to the methanol slurry and the slurry was shaken until all non- membrane contents of the bacteria are transferred to the methanol-acetone fraction. The methanol and acetone were then evaporated to leave a dry crude extract. The extract was stored in the dark for long-term use. Prior to use, the dry crude extract was resuspended in isopropanol. Upon resuspension, the membrane components typically settled to the bottom of the container and were separated from the supernatant. However, a fraction of the membrane components (about 15%) did remain in the supernatant.

[0102] The inhibitory dose of different extracts was determined by a growth assay as follows: different volumes (0.5%, 1% and 2% (v / v) doses) of the extracts were added to 2 mL potato dextrose broth or potato dextrose agar and inoculated ~102Aspergillus flavus spores in the medium. An untreated control was also included. Triplicate samples were grown in each group (control as well as untreated) for 72 hours at 30°C and colony diameters recorded. Any treatment that reduced colony diameter by >50% was inhibitory. It was noticed that treatments below 1% were not inhibitory. Depending on the strain, adjustments of the the dose but, in general, it was to be >=1%.Attorney Docket No.1655.3.WO Example 3: Broad Spectrum Antimicrobial Activity

[0103] To demonstrate the antimicrobial activity of Vibrio gazogenes, ATCC Accession No. 43943, different amounts of extract were tested against a panel of human pathogens (Table 1) and agriculturally relevant fungal pathogens (FIG. 5). This analysis indicated that the Vibrio gazogenes, ATCC Accession No.43943, extract exhibited dose-dependent inhibition of the growth of human pathogens Porphyromonas gingivalis, Enterococcus faecium, Candida sp. and Aspergillus fumigatus as well as plant pathogenic fungi Rhizoctonia solani, Penicillium expansum, Botrytis cinerea, Verticillium dahlia, Fusarium oxysporum, and Phoma betae. Table 1. Antimicobial activity against human pathogens Inhibition zone on solid agar growth medium after 3 days (mm) Human PathogenDose of treatment with ATCC 43943 Extract

[0104] Therefore, the antimicrobial extract of this invention finds use in a number of applications. For example, the extract may be used as a biocontrol agent against plant pathogens, including fungi and bacteria that cause crop diseases. This can be accomplished by applying the extract to the soil or plant surfaces, or by incorporating the extract into plant growth media. Additionally, the extract may be used as a seed treatment to protect the seeds from microbial infections. The extract may also be applied to crops during their growth cycle to protect against microbial infections, resulting in higher yields and better crop quality.

[0105] The extract of Vibrio gazogenes of the present invention may have potential applications in the health industry as well. For example, it could be used as an active ingredient in topical creams, ointments, or sprays for treating skin infections caused by bacteria or fungi. The extract may also be incorporated into disinfectant solutions for use in hospitals, clinics, and other healthcare facilities. The extract may also have potential as a natural preservative for food and cosmetic products, due to its antimicrobial properties.Furthermore, the antimicrobial extract may be used in the production of agricultural products, such as crops and livestock. The extract may beAttorney Docket No.1655.3.WO added to animal feed to prevent the growth of harmful bacteria, fungi, and other microbes, thereby improving the health and productivity of the animals. Example 4: Compositional Analysis of Vibrio gazogenes, ATCC Accession No.43943, Extract

[0107] To determine the compositional components of the extract, untargeted metabolomics analysis using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and proteome analysis (using NanoLC-MS / MS Analysis) was performed on triplicate samples. The material tested was a dried extract prepared as described in Example 2. For proteomic analysis, the dried extract was suspended in methanol before injecting in UPLC (metabolomics) or NanoLC (proteome). As presented in Table 2, the composition contains a plurality of bioactive metabolites. Table 2. Composition of Vibrio gazogenes, ATCC Accession No.43943, Extract Constituent Percent* Property Antibacterial, anticancer, andarde ce p eva e ce o a y o e e a o e s ess a . . Example 5: Seed Coating Application in Corn

[0108] Field studies with a seed coating application were conducted. For seed treatment application, we used a formulation consisting of 1:6 methanol extract: secondary solvent formulation (consisting of 80% isopropanol and 20% propylene glycol and we diluted this with 100X water. The seeds were treated at an application rate of 15 mL with this formulation per 100000 corn seeds. The results of this analysis indicated that seed treatment reduced fungalAttorney Docket No.1655.3.WO infections in corn seeds, promoted germination, growth rate (FIG.6A) and overall health (FIG. 6B) and biomass production (FIG.6C) of corn seedlings. Example 6: Applications in Sugar Beet

[0109] We have also tested the potential of the formulation to prevent Rhizoctonia root rot in artificially infected sugar beet roots, in which co-infection was done by inoculating Rhizoctonia solani (a fungal pathogen) and Leuconostoc mesenteroides (a bacterial pathogen). The sugar beet cultivar B-7 roots were inoculated on the shoulder of the root (where root meets the soil line) using a cork borer to create a 10-mm diameter × 24-mm deep hole. For the non-inoculated control, 0.1 mL of the vector (formulation without the extract) was inserted. L. mesenteroides artificial infection was initiated by inoculating 0.1 mL of a 108cfu / mL bacterial suspension. R. solani infection was initiated by placing a 2 × 2 mm piece of mycelial mass in the hole with 0.1 mL of sterile well water. For the treatment application, we used a formulation consisting of 1:6 methanol extract: secondary solvent formulation (consisting of 60% isopropanol and 40% propylene glycol and we diluted this with 100X water. A 0.1 mL of this formulation was applied to the plants. Post inoculation, the plugs were replaced and sealed with petroleum jelly.

[0110] The roots were evaluated for rot by bisecting the root through the inoculation hole and measuring the amount of rot perpendicular to the plug with a ruler. The data presented in Table 3 represent mean ± Standard Error of ten biological replicates. Table 3. Prevention of Root Rot in Sugar Beet Sample group Root rot (mm) ance waseterm ne w t tu ents t-test (p . cons ere s gn cant; n= ).

[0111] In the sugar beet crop, significant increases in biomass and sucrose content per biomass (Table 4) were observed. Table 4. Increases in Sucrose ContentAttorney Docket No.1655.3.WO Sample group Sucrose content (µg / mg) Samples with infection and no treatment 195 ± 7 ance was

[0112] We have also tested the potential of the formulation to prevent disease symptoms caused by artificial leaf infection of a sugar beet crop with Cercospora beticola. For the foliar spray treatment application, we used a formulation consisting of 1:6 methanol extract: secondary solvent formulation (consisting of 60% isopropanol and 40% propylene glycol and we diluted this with 100X water. Cercospora leaf spot disease rating was assigned based on established methods. Briefly, Cercospora leaf spot severity was rated on the leaf spot assessment scale of 1 to 10 (Jones & Windels (1991) Minnesota Extension Service. University of Minnesota. AG-FO-5643-E). A rating of 1 indicated the presence of 1-5 spots / leaf or 0.1% disease severity, and a rating of 10 indicated 50% or higher disease severity. This analysis indicated that foliar spray reduced Cercospora infections in sugar beet leaves (Table 5). Table 5. Decreases in Cercospora infection Treatment Disease Rating ance was- p . g ; .

[0113] The foregoing is illustrative of the present invention and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.

Claims

Attorney Docket No.1655.3.WO WHAT IS CLAIMED IS:

1. A method of producing an antimicrobial extract comprising (a) contacting cells of Vibrio gazogenes, ATCC Accession No.43943, with methanol; and (b) extracting the cells of (a) with one or more of acetone, chloroform, acetonitrile, isopropanol, propylene glycol, or dimethylsulfoxide thereby producing the antimicrobial extract, the extract being substantially devoid of membrane components and comprising compounds soluble in (i) methanol and (ii) acetone, chloroform, acetonitrile, isopropanol, propylene glycol, and / or dimethylsulfoxide, thereby producing the antimicrobial extract.

2. The method of claim 1, wherein prior to extracting, the method further comprises cultivating the Vibrio gazogenes in a culture medium and separating the Vibrio gazogenes cells from the medium.

3. The method of claim 2, wherein cultivating the Vibrio gazogenes comprises cultivating the Vibrio gazogenes in marine broth medium at about 26°C for about 18 hours to 24 hours.

4. The method of any one of claims 1-3, further comprising evaporating the (a) methanol and (b) acetone, chloroform, acetonitrile, isopropanol, propylene glycol, and / or dimethylsulfoxide to produce a dried anti-microbial extract.

5. The method of claim 4, further comprising resuspending the dried anti-microbial extract in methanol, ethanol, isopropanol, chloroform, acetonitrile, ether, propylene glycol, and / or dimethylsulfoxide.

6. An antimicrobial extract of Vibrio gazogenes produced by the method of any one of claims 1-5.

7. The antimicrobial extract of claim 6, said antimicrobial extract comprising, based upon the total, about 30% to about 32% prodigiosin, about 3% to about 4% 6-hydroxycaproicAttorney Docket No.1655.3.WO acid, about 1% to about 2% acetophenone, about 0.5% to about 1% 2'-deoxyadenosine, about 0.01% to about 0.05% baquiloprim, and about 5% to about 30% peptide.

8. A commodity, industrial, or commercial product comprising the extract of claim 6 or 7.

9. The product of claim 8, wherein the product is (a) a pharmaceutical composition suitable for administration orally, dermally, parenterally, nasally, ophthalmically, otically, sublingually, rectally and / or vaginally; (b) a food product including a fish, crustacean, fish substitute, crustacean substitute, meat, meat substitute, poultry product, vegetable, green, sauce, emulsion, beverage, juice, wine, beer, dairy product, egg-based product, jam, jelly, grain-based product, baked good or confectionary product; (c) a cosmetic or personal care product including a cream, gel, powder, lotion, sunscreen, lipstick, body wash, herbal extract, or formulation that supports the growth of bacteria; (d) an agricultural product including a foliar spray, root drench, seed coating, or animal feed; or (e) a surface cleaning product including a cleaning product for counter tops, doors, windows, handles, surgical equipment, or medical tools.

10. A method for preventing or reducing the growth of a microbial pathogen comprising contacting a microbial pathogen with an effective amount of the antimicrobial extract of claim 6 or 7, thereby preventing or reducing the growth of the microbial pathogen.

11. The method of claim 10, wherein the microbial pathogen is a fungal pathogen comprising Aspergillus flavus, Aspergillus parasiticus, Aspergillus fumigatus, Rhizoctonia solani, Candida albicans, Podosphaera aphanis, Penicillium expansum, Botrytis cinerea, Verticillium dahlia, Fusarium oxysporum, Phoma betae, Cersospora sp, Magnaporthe sp. or Phyllachora sp.Attorney Docket No.1655.3.WO 12. The method of claim 10, wherein the microbial pathogen is a bacterial pathogen comprising Leuconostoc mesenteroides, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Porphyromonas gingivalis, Enterococcus faecium, Salmonella sp. or Listeria monocytogenes.

13. A method for preventing or reducing the likelihood of contamination of a food product by a microbial pathogen comprising applying an effective amount of the antimicrobial extract of claim 6 or 7 to the food product, thereby preventing or reducing the likelihood of contamination of the food product with a microbial pathogen.

14. The method of claim 13, wherein the food product is a beverage, fruit or vegetable.

15. A method for inhibiting the growth of a microbial pathogen on an inanimate surface or environmental sample comprising contacting an inanimate surface or environmental sample with an effective amount of the antimicrobial extract of claim 6 or 7, thereby inhibiting the growth of a microbial pathogen on the inanimate surface or environmental sample.

16. The method of claim 15, wherein growth of the microbial pathogen is inhibited by about 50% to about 90% as compared to a control.

17. The method of claim 15, wherein the inanimate surface is building material.

18. A method for preventing, treating, or reducing the likelihood of infection by a microbial pathogen comprising administering to a subject in need thereof an effective amount of the antimicrobial extract of claim 6 or 7, thereby preventing, treating, or reducing the likelihood of infection by the microbial pathogen.

19. The method of claim 18, wherein the microbial pathogen is a bacterium or a fungus.Attorney Docket No.1655.3.WO 20. The method of claim 18, wherein the bacterium is Escherichia coli, Salmonella sp., Listeria sp., Staphylococcus aureus, Pseudomonas areuginosa, Enterococcus faecium, Porphyromonas gingivalis.

21. The method of claim 18, wherein the fungus is an Aspergillus sp. or Candida sp.

22. A method of treating, preventing or reducing the likelihood of infection of a plant or part thereof by a microbial pathogen comprising contacting the plant or part thereof with an effective amount of the antimicrobial extract of claim 6 or 7, thereby treating, preventing, or reducing the likelihood of infection by the microbial pathogen of the plant or part thereof.

23. The method of claim 22, wherein the microbial pathogen is a bacterium or a fungus.

24. The method of claim 22, wherein the bacterium is Pseudomonas aeruginosa.

25. The method of claim 22, wherein the fungus is Aspergillus flavus, Aspergillus parasiticus, Aspergillus fumigatus, Rhizoctonia solani, Podosphaera aphanis, Penicillium expansum, Botrytis cinerea, Verticillium dahlia, Fusarium oxysporum, Phoma betae, Cersospora sp, Magnaporthe sp. or Phyllachora sp.

26. An antimicrobial extract of Vibrio gazogenes, ATCC Accession No.43943, the extract being substantially devoid of membrane components and comprising compounds soluble in (a) methanol and (b) acetone, ethanol, isopropanol, chloroform, acetonitrile, or dimethylsulfoxide, wherein the compounds comprise prodigiosin, 6-hydroxycaproic acid, acetophenone, 2'-deoxyadenosine, baquiloprim and peptides.

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