Gas phase treatment of agricultural products

Abstract

A method of treatment of agricultural products that mitigates potential infections such as fungal infections and delays maturation of agricultural products is described. The agricultural product treatment method includes providing an antimicrobial agent to a shell comprising a plurality of agricultural product articles and contacting the plurality of agricultural product articles with the antimicrobial agent wherein the antimicrobial agent is in gaseous form and is selected to inactivate latent microorganisms in the plurality of agricultural product articles.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. application serial number 63 / 322,176, filed on March 21, 2022, entitled “GAS PHASE TREATMENT OF LATENT INFECTION IN PRODUCE,” and U.S. application serial number 63 / 486,892, filed on February 24, 2023, entitled “GAS PHASE TREATMENT OF PRODUCE,” which are incorporated herein by reference in their entirety. Technical Field

[0003] The present invention relates to the use of a slow-release, gas phase treatment process to alleviate latent infections, such as fungal infections, and to delay ripening of produce. In addition, the present invention relates to an edible coating composition comprising one or more monoglycerides, one or more fatty acid salts, and allyl isothiocyanate. Background Art

[0004] Post-harvest food spoilage is a significant factor limiting the storage and market life of agricultural products, resulting in significant economic losses worldwide. The main causes of these losses are insect infestations or diseases or improper storage conditions, which lead to decay or fresh weight loss due to respiration and evaporation. Unlike other more common post-harvest fungal infections that require wounds to enter and cause decay, latent infections originate before the produce is harvested and reside within the produce. Latent infections are generally not prevented or mitigated by common surface treatments used for other plant pathogens that cause decay.

[0005] In addition, many common agricultural products, such as avocados and bananas, are often harvested before they are fully ripe and then allowed to fully ripen after harvest, such as during storage or transportation. Many of these products are seasonal and therefore can only ripen within a limited time window. This often limits consumers to only being able to obtain the seasonal agricultural products within that limited time window. SUMMARY OF THE INVENTION

[0007] The present invention describes the use of slow-release gas phase treatment methods to alleviate latent fungal infections and delay maturity. In some embodiments, biofumigation as described herein uses molecules present in fruits and vegetables as a natural method for managing agricultural products after harvest diseases. Concentrated forms have reactive or toxic natural compounds that may be harmful to transportation and processing, and are difficult to store. These active compounds are stored with stable precursors and converted into active forms enzymatically on the spot will simplify transportation and processing, and extend shelf life. Enzymatic processes are carried out at high rates under mild conditions (close to ambient temperature, atmospheric pressure and physiological pH value). Enzymatic processes are more environmentally friendly, more cost-effective, and ultimately more sustainable.

[0008] While the disclosed inventive concepts include those defined in the appended claims, it should be understood that these inventive concepts may also be defined in terms of the following embodiments.

[0009] In addition to the embodiments of the appended claims and the embodiments described above, the following numbered embodiments are also inventive.

[0010] Embodiment 1 includes a method for processing agricultural products, comprising:

[0011] providing an antimicrobial agent to the enclosure; and

[0012] A plurality of produce items are contacted with the antimicrobial agent, wherein the antimicrobial agent is in gaseous form and is selected to inactivate latent microorganisms in the plurality of produce items.

[0013] Embodiment 2 is a method according to embodiment 1, further comprising sealing the housing before providing the antimicrobial agent to the housing.

[0014] Embodiment 3 is a method according to embodiment 1 or 2, wherein the housing comprises an agricultural product packaging container, an agricultural product storage container, an agricultural product transport container, or an agricultural product handling container.

[0015] Embodiment 4 is a method according to embodiment 3, wherein the produce packaging container comprises a clamshell.

[0016] Embodiment 5 is a method according to embodiment 3, wherein the produce transport container comprises a shipping container or a box truck.

[0017] Embodiment 6 is a method according to embodiment 3, wherein the produce processing vessel comprises a chlorosis chamber or a ripening chamber.

[0018] Embodiment 7 is a method according to any one of embodiments 2-6, wherein the housing is sealed or the diffusion rate of gas in and out of the housing is restricted.

[0019] Embodiment 8 is a method according to any one of embodiments 1-7, wherein providing the antimicrobial agent to the housing comprises providing a gas stream containing the antimicrobial agent to the housing.

[0020] Embodiment 9 is a method according to embodiment 8, further comprising pumping the gas flow into the housing.

[0021] Embodiment 10 is the method of any one of embodiments 1-9, wherein the length of time is in the range of 1 minute to 100 hours.

[0022] Embodiment 11 is a method according to embodiment 10, wherein the length of time is in the range of 10 minutes to 1 hour.

[0023] Embodiment 12 is a method according to any one of embodiments 1-11, further comprising adjusting the temperature or humidity inside the housing.

[0024] Embodiment 13 is a method according to any one of embodiments 1-12, further comprising adjusting the temperature or humidity inside the housing.

[0025] Embodiment 14 is a method according to embodiment 13, wherein volatilization of the antimicrobial agent precursor occurs outside of the housing.

[0026] Embodiment 15 is a method according to embodiment 13, wherein volatilizing the antimicrobial agent precursor occurs within the shell.

[0027] Embodiment 16 is a method according to any one of embodiments 1-15, further comprising activating an antimicrobial agent precursor to produce the antimicrobial agent.

[0028] Embodiment 17 is a method according to embodiment 16, wherein activating the antimicrobial agent precursor comprises contacting the antimicrobial agent precursor with a catalyst selected to promote conversion of the antimicrobial agent precursor to the antimicrobial agent.

[0029] Embodiment 18 is a method according to embodiment 17, wherein the catalyst is a biocatalyst.

[0030] Embodiment 19 is a method according to embodiment 18, wherein the biocatalyst is an enzyme.

[0031] Embodiment 20 is a method according to embodiment 19, wherein the antimicrobial agent precursor comprises alliin, the enzyme comprises alliinase, and the antimicrobial agent comprises allicin.

[0032] Embodiment 21 is a method according to embodiment 19, wherein the antimicrobial agent comprises sinigrin, the enzyme comprises myrosinase, and the antimicrobial agent comprises allyl isothiocyanate.

[0033] Embodiment 22 is the method of any one of embodiments 1-21, wherein providing the antimicrobial agent to the housing comprises:

[0034] providing an antimicrobial precursor to a reactor, wherein the reactor comprises a catalyst selected to promote conversion of the antimicrobial precursor to the antimicrobial agent;

[0035] flowing a carrier gas from the first end of the reactor to the second end of the reactor to form a gaseous mixture comprising the carrier gas and the antimicrobial agent; and directing the gaseous mixture from the second end of the fixed bed reactor to the housing.

[0036] Embodiment 23 is a method according to embodiment 22, wherein providing the antimicrobial agent precursor to the reactor comprises pumping the antimicrobial agent precursor to the reactor.

[0037] Embodiment 24 is a method according to embodiment 23, wherein flowing the gas from the first end of the reactor to the second end of the reactor includes forcing the gas to flow from the first end of the reactor to the second end of the reactor.

[0038] Embodiment 25 is a method according to embodiment 24, wherein the reactor is a fixed bed reactor.

[0039] Embodiment 26 is a method according to embodiment 24, wherein the catalyst comprises an enzyme.

[0040] Embodiment 27 is a method according to any one of embodiments 1-26, wherein contacting the plurality of produce items with the antimicrobial agent comprises circulating the antimicrobial agent in the housing.

[0041] Embodiment 28 is a method according to any one of embodiments 1-27, wherein contacting the plurality of produce items with the antimicrobial agent comprises penetrating a skin layer of each of the plurality of produce items with the antimicrobial agent.

[0042] Embodiment 29 is a method according to any one of embodiments 1-28, wherein the antimicrobial agent comprises one or more of an antimicrobial agent, an antifungal agent, and an antiviral agent.

[0043] Embodiment 30 is a method according to embodiment 29, wherein the antimicrobial agent comprises an antifungal agent.

[0044] Embodiment 31 is a method according to embodiment 30, wherein the antifungal agent comprises allyl isothiocyanate, diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranilate, methyl cinnamate, γ-decalactone, α-terpineol, linalool, or any combination thereof.

[0045] Embodiment 32 is a method according to embodiment 30, wherein the antifungal agent comprises a derivative of cis-3-hexene.

[0046] Embodiment 33 is a method according to embodiment 32, wherein the antifungal agent comprises cis-3-hexenol or cis-3-hexenyl acetate.

[0047] Embodiment 34 is a method according to any one of embodiments 30-33, wherein the antifungal agent is selected to inactivate green mold or penicillium mold.

[0048] Embodiment 35 is a method according to any one of embodiments 30-34, wherein the antifungal agent is selected to inactivate Botrytis cinerea, Penicillium spp., Monilinia spp., Alternaria alternata, Rhizopus stolonifera, Trichothecium roseum, Fusarium spp., Colletotrichum spp., or any combination thereof.

[0049] Embodiment 36 is a method according to any one of embodiments 1-35, wherein the plurality of produce items comprises fruits, vegetables, or a combination thereof.

[0050] Embodiment 37 is a method according to embodiment 36, wherein the plurality of produce items includes fruit.

[0051] Embodiment 38 is a method according to embodiment 37, wherein the fruit comprises berries (e.g., avocado, banana, blueberry, cranberry, eggplant, tomato, grape, persimmon), citrus fruits (e.g., orange, lemon, lime, grapefruit, kumquat), gourd fruits (e.g., pumpkin, cucumber, watermelon), stone fruits (e.g., peach, plum, cherry, olive), pome fruits (e.g., apple, pear), aggregate fruits (e.g., blackberry, raspberry), and accessory fruits (e.g., strawberry).

[0052] Embodiment 39 is a method according to any one of embodiments 1-38, further comprising placing the plurality of produce items in the enclosure prior to providing the antimicrobial agent to the enclosure.

[0053] Embodiment 40 is a method according to any one of embodiments 1-39, further comprising, after contacting the plurality of produce items with the antimicrobial agent for a period of time, removing the plurality of produce items from the housing to obtain a plurality of treated produce items.

[0054] Embodiment 41 is a method according to any one of embodiments 1-40, wherein the antimicrobial agent comprises allyl isothiocyanate.

[0055] Embodiment 42 is a method according to any one of embodiments 1-41, wherein the antimicrobial agent comprises allyl isothiocyanate and one or more antimicrobial agents selected from the group consisting of:

[0056] Diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranilate, methyl cinnamate, gamma-decanolide, alpha-terpineol, and linalool.

[0057] Embodiment 43 is a method according to embodiment 41 or 42, wherein the antimicrobial agent is provided to the housing at a concentration of at least about at least 0.1 ppmv, at least 1 ppmv, at least 10 ppmv, at least 100 ppmv, at least 1000 ppmv, at least about 1200 ppmv, at least about 1250 ppmv, or about 0.1 ppmv to about 4900 ppmv, or about 1250 ppmv to about 5000 ppmv, or about 1250 ppmv to about 4900 ppmv.

[0058] Embodiment 44 is a method according to embodiment 38, wherein the fruit is avocado, strawberry, citrus or lemon.

[0059] Embodiment 45 is the method of any one of embodiments 1-44, further comprising coating the plurality of produce items with an edible coating composition.

[0060] Embodiment 46 is a method according to embodiment 45, wherein the edible coating composition comprises:

[0061] a coating agent comprising one or more saturated glycerides selected from monoglycerides and diglycerides; and one or more fatty acid salts; and

[0062] Solvent.

[0063] Embodiment 47 is a method according to embodiment 46, wherein the one or more saturated glycerides are present in the coating agent in an amount of about 75 wt% to about 98 wt%, or about 80 wt% to about 98 wt%, or 85 wt% to about 98 wt%, or about 90 wt% to about 98 wt%, or about 92 wt% to about 97 wt%, or about 95 wt%.

[0064] Embodiment 48 is a method according to embodiment 46 or 47, wherein the one or more fatty acid salts are present in the coating agent in an amount of about 1 wt% to about 20 wt%, or about 1 wt% to about 15 wt%, or about 1 wt% to about 12 wt%, 2 wt% to about 8 wt%, or about 3 wt% to about 7 wt%, or about 5 wt%.

[0065] Embodiment 49 is a method according to any one of embodiments 46-48, wherein each of the one or more saturated monoglycerides comprises a carbon chain length of about C10 to about C22.

[0066] Embodiment 50 is a method according to any one of embodiments 46-49, wherein one of the one or more saturated glyceryl monoesters is glyceryl monostearate.

[0067] Embodiment 51 is a method according to any one of embodiments 46-50, wherein each of the one or more fatty acid salts comprises a carbon chain length of about C10 to about C22.

[0068] Embodiment 52 is a method according to any one of embodiments 46-51, wherein one of the fatty acid salts is sodium stearate.

[0069] Embodiment 53 is a method according to any one of embodiments 46-52, wherein the coating agent further comprises one or more antimicrobial agents, wherein the one or more antimicrobial agents are selected from diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranil, methyl cinnamate, γ-decanoic acid, α-terpineol, linalool, cis-3-hexenol or cis-3-hexenyl acetate, sodium benzoate, potassium sorbate, chalcone, fludioxonil, 2-hydroxychalcone ketone, 4-hydroxychalcone, 4'-hydroxychalcone, 2,2'-dihydroxychalcone, 2,4'-dihydroxychalcone, 2',4-dihydroxychalcone, 2',4'-dihydroxychalcone, 2',4,4'-trihydroxychalcone, 2',4,4'-trihydroxychalcone intermediate, violastyrene, obtusaquinone, apiole, piperine, celastrol, arthonoic acid acid, leoidin, antimycin A, antimycin A1, refractory lichenic acid, ethylorsellinate, methyl orsellinate, mycophenolic acid, ethyldichloroorsellinate, angoraxin, isocarpone, eupatoriochromene, xanthoxylum, lichenic acid, aloesin, formononetin, acetovanillin, isoorsenflavone, deoxysulphurone B 7,4'-dimethyl ether, chrysin dimethyl ether, bergamotol, gambogic acid, 2-hydroxyxanthenone, isoanisole, zanthoxylum lactone, acetylhymetochrome, nobiletin, hymechrome, methoxsalen, 4-methylesculetin, tangerine, kaelin, flavonoids, 3,4',5,6,7-pentamethoxyflavone, deguelin (-), limonene, deoxysappanone B trimethyl ether, deoxysappanone B 7,3'-dimethyl ether, 2',4'-dihydroxy-4-methoxychalcone, daunorubicin hydrochloride, plumbagin, menadione, musciquinone, levomenthol, methyl trimethoxycinnamate, chavicol, cinnamylphenol, benzoate, napthoquinone, benzophenone, acetophenone, benzophenone, phenylacetophenone, chitosan, salicylic acid and sodium salicylate.

[0070] Embodiment 54 is an agricultural product processing system, comprising:

[0071] A reactor configured to facilitate conversion of an antimicrobial agent precursor to an antimicrobial agent, wherein the reactor comprises:

[0072] a bed configured to contain a particulate media coupled to a catalyst, wherein the catalyst is selected to promote conversion of the antimicrobial agent precursor to the antimicrobial agent;

[0073] an inlet proximate a first end of the bed and configured to direct a carrier gas from the first end of the bed to a second end of the bed;

[0074] an additional inlet configured to direct the antimicrobial agent precursor to the bed; and

[0075] an outlet proximate a second end of the bed and configured to allow a gas mixture comprising the carrier gas and the antimicrobial agent to exit; and

[0076] A housing is fluidly coupled to the reactor via the outlet, wherein the housing is configured to receive a plurality of produce items and contain a gas mixture in contact with the plurality of produce items.

[0077] Embodiment 55 is an agricultural product processing system according to embodiment 54, wherein the reactor comprises a fixed bed reactor.

[0078] Embodiment 56 is the agricultural produce processing system of embodiment 54 or 55, further comprising a pump in fluid communication with the additional inlet and configured to provide the antimicrobial precursor to the reactor.

[0079] Embodiment 57 is an agricultural product processing system according to embodiment 56, wherein the pump is a peristaltic pump.

[0080] Embodiment 58 is a produce processing system according to any one of embodiments 54-57, wherein the carrier gas provided to the inlet volatilizes the antimicrobial agent precursor.

[0081] Embodiment 59 is a plurality of produce items treated using the method of any one of embodiments 1-53.

[0082] Embodiment 60 is a plurality of produce items according to embodiment 59, wherein the plurality of produce items have a mass loss factor of at least 1.50, at least 1.6, at least 1.7, at least 1.8, at least 1.9, at least 2.0, or at least 2.5.

[0083] Embodiment 61 is a plurality of produce items according to embodiment 59 or 60, wherein the plurality of produce items are avocados, strawberries, citrus, or lemons.

[0084] Embodiment 62 is a plurality of produce items according to embodiment 61, wherein the plurality of produce items are 30 treated California #60 avocados, and 5 days after the 30 treated California #60 avocados are treated, the average Shore value of the 30 treated California #60 avocados is at least 10 Shores, at least 15 Shores, at least 20 Shores, at least 25 Shores higher than the average Shore value of 30 untreated California #60 avocados, and wherein at day 0, the 30 treated California #60 avocados have an average Shore value that is ±10 Shores of the 30 untreated California #60 avocados.

[0085] Embodiment 63 is a method for improving the shelf life of multiple agricultural products, the method comprising: providing an antimicrobial agent to a shell containing multiple agricultural products; and contacting the multiple agricultural products with the antimicrobial agent, wherein the antimicrobial agent is in gaseous form and is selected to inactivate latent microorganisms in the multiple agricultural products.

[0086] Embodiment 64 is a method according to embodiment 63, further comprising coating the plurality of produce items with an edible coating composition.

[0087] Embodiment 65 is a method for delaying the ripening of multiple agricultural products, the method comprising: providing an antimicrobial agent to a shell containing multiple agricultural products; and contacting the multiple agricultural products with the antimicrobial agent, wherein the antimicrobial agent is in gaseous form and is selected to inactivate latent microorganisms in the multiple agricultural products.

[0088] Embodiment 66 is a method according to embodiment 65, further comprising coating the plurality of produce items with an edible coating composition.

[0089] Embodiment 67 is an edible coating composition comprising: a coating agent comprising: one or more monoglycerides, one or more fatty acid salts and allyl isothiocyanate; and a solvent.

[0090] Embodiment 68 is a composition according to embodiment 67, wherein the allyl isothiocyanate is present in the edible coating composition in an amount from about 0.001% v / v to about 5% v / v, or from about 0.01% v / v to about 2.5% v / v, or from about 0.1% v / v to about 1.5% v / v, or from about 0.5% v / v to about 1.5% v / v.

[0091] Embodiment 69 is a composition according to embodiment 67 or 68, wherein the one or more monoglycerides are present in the coating agent in an amount of about 75 wt% to about 98 wt%, about 80 wt% to about 98 wt%, or 85 wt% to about 98 wt%, or about 90 wt% to about 98 wt%, or about 92 wt% to about 97 wt%, or about 95 wt%.

[0092] Embodiment 70 is a composition according to any of embodiments 67-69, wherein the one or more fatty acid salts are present in the coating agent in an amount of about 1 wt % to about 20 wt %, or about 1 wt % to about 15 wt %, or about 1 wt % to about 12 wt %, 2 wt % to about 8 wt %, or about 3 wt % to about 7 wt %, or about 5 wt %.

[0093] Embodiment 71 is a composition according to any one of embodiments 67-70, wherein each of the one or more monoglycerides comprises a carbon chain length of about C10 to about C22.

[0094] Embodiment 72 is a composition according to any one of embodiments 67-71, wherein one of the one or more saturated monoglycerides is glyceryl monostearate.

[0095] Embodiment 73 is a composition according to any one of embodiments 67-72, wherein each of the one or more fatty acid salts comprises a carbon chain length of about C10 to about C20.

[0096] Embodiment 74 is a composition according to any one of embodiments 67-73, wherein one of the fatty acid salts is sodium stearate.

[0097] Embodiment 75 is a composition according to any one of embodiments 67-74, wherein the solvent comprises water.

[0098] Embodiment 76 is a composition according to any of embodiments 67-75, wherein the coating agent is present in an amount ranging from about 1 g / L to about 150 g / L, or from about 10 g / L to about 100 g / L, or from about 25 g / L to about 75 g / L, or from about 40 g / L to about 60 g / L, or from about 45 g / L to about 55 g / L.

[0099] Embodiment 77 is a plurality of produce items coated with the edible coating composition of any one of embodiments 67-76.

[0100] Embodiment 78 is a plurality of agricultural produce items according to Embodiment 77, wherein the plurality of agricultural produce items have a mass loss factor of at least 1.50, at least 1.6, at least 1.7, at least 1.8, or at least 2, or at least 2.5.

[0101] Embodiment 79 is a plurality of produce items according to embodiment 77 or 78, wherein the plurality of produce items are avocados.

[0102] As used herein, the term "antimicrobial" refers to compounds that inhibit the growth of microorganisms, including inhibiting the growth of bacteria, fungi, and viruses.

[0103] As used herein, "fatty acid" is a hydrocarbon chain containing an ester, acid or carboxylate group, which is collectively referred to as an "oxycarbonyl moiety" and is bonded to one end of the hydrocarbon chain, which is understood to be the "hydrophilic" end; while the opposite end is understood to be the "hydrophobic" end. Fatty acids include fatty acids, fatty acid esters (e.g., monoglycerides, diglycerides), and fatty acid salts. The term "glyceride" used herein refers to an ester formed by glycerol and at least one fatty acid, which is a fatty acid derivative.

[0104] As used herein, a glycerol monoester having a carbon chain length of, for example, about C10 to about C22 means that the lipophilic portion of the glycerol monoester has from about 10 carbon atoms to about 22 carbon atoms. For example, a C18 glycerol monoester would include glycerol monostearate, since the lipophilic portion of the molecule (e.g., monostearate) has a carbon chain length of 18 carbon atoms, e.g., (C 18 H 35 O)-.

[0105] As used herein, "glyceryl" refers to a propyl group in which each of the two carbon atoms not centered on the group is substituted with a hydroxyl group. In some embodiments, the glyceryl group is a 1-glyceryl group (i.e., -CH2CH(OH)CH2OH). In some embodiments, the glyceryl group is a 2-glyceryl group (i.e., -CH(CH2OH)CH2OH).

[0106] The term "mass loss rate" refers to the rate at which the produce loses mass (eg, by releasing water and other volatile compounds). The mass loss rate is typically expressed as a percentage of the original mass per unit time (eg, a percentage per day).

[0107] The term "mass loss factor" refers to the ratio of the average mass loss rate of uncoated agricultural products (measured for the control group) to the average mass loss rate of the corresponding tested agricultural products (e.g., coated agricultural products) over a given period of time. Therefore, a larger mass loss factor of the coated agricultural product corresponds to a greater reduction in the average mass loss rate of the coated agricultural product.

[0108] The term "weight %" or "weight percent" refers to the percentage represented by the identified component or components, which percentage is calculated as the weight percent of all components excluding water, unless otherwise indicated.

[0109] The details of one or more embodiments of the subject matter of the present invention are shown in the drawings and the specification. Other features, aspects and advantages of these subjects can be apparent from the specification, drawings and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0110] Figure 1 is a flow chart showing the steps in one embodiment of a method of processing produce.

[0111] Figure 2 is a schematic diagram showing one embodiment of a system for gas phase treatment of latent infections in produce.

[0112] Figure 3 It is a graph showing the incidence of infection in citrus. The antifungal activity of diallyl disulfide (DADS) vapor against Penicillium digitatum (Pd) was tested.

[0113] Figure 4 is a graph of the incidence of infection in citrus. The antifungal activities of allyl isothiocyanate (AITC) and DADS vapor against Pd and Penicillium italicum (Pi) were tested.

[0114] Figure 5 is a graph of infection incidence in citrus, where the antifungal activity of low dose AITC vapor against Pd was tested.

[0115] Figure 6 is a graph of the incidence of infection in citrus, where a range of AITC vapor concentrations were tested for antifungal activity against Pd.

[0116] Figure 7 is a graph of infection incidence in citrus, where a range of AITC vapor exposure times were tested for the antifungal activity of Pd.

[0117] Figure 8 is a graph of infection incidence in citrus, testing the antifungal activity of AITC vapor against Pd after a range of pre-treatment incubation times.

[0118] Fig. 9 is a graph of infection incidence in avocados, the antifungal activity of AITC and DADS vapors against Colletotrichum gloeosporioides (Cg) was tested.

[0119] Fig.10 is a graph of infection incidence in avocados tested for antifungal activity of Cg over a range of DADS vapor exposure times.

[0120] Fig.11 is a graph of the incidence of infection in avocados, where a range of AITC and carvacrol vapor exposure times were tested for the antifungal activity of Cg.

[0121] Fig.12 is a graph of the Shore hardness results for avocados, testing the antifungal activity of Cg over a range of AITC and carvacrol vapor exposure times.

[0122] Fig.13 is a graph of the incidence of infection in avocados tested for the antifungal activity of Cg over a range of AITC vapor exposure times.

[0123] Fig.14 is a graph of durometer shore results for avocados tested for antifungal activity of Cg over a range of AITC vapor exposure times.

[0124] Fig.15 is an image of an avocado half exposed to 500 μL of AITC vapor.

[0125] Fig.16 Schematic representation of the enzyme pathways hypothesized to be associated with thickening of avocado peel after exposure to AITC vapor.

[0126] Fig.17 is a graph showing the incidence of infection in grapes. The antifungal activity of AITC and DADS vapors against Botrytis cinerea (Bc) was tested.

[0127] Fig.18 is a graph of the infection incidence in strawberry, where the antifungal activity of AITC vapor was tested against Bc.

[0128] Fig.19 is a graph showing the incidence of infection in two test groups of strawberries, tested for antifungal activity against Bc over a range of AITC vapor concentrations.

[0129] Fig. 20 is a graph of the infection incidence of strawberry, and the antifungal activity of AITC vapor against Bc was examined under different exposure conditions.

[0130] Fig.21 is a graph showing the incidence of infection in strawberries treated with the coating agents, and the antifungal activity of AITC vapor against Bc was tested.

[0131] Fig. 22is a graph showing the results of mass loss factor (MLF) of strawberries hung and treated with coating agents, and the antifungal activity of AITC against Bc was tested.

[0132] Fig.23 is a graph of the percentage of mold in strawberries hung and treated with coating agents, and the antifungal activity of AITC against Bc was tested.

[0133] Fig.24 is a graph of infection incidence in citrus. The antifungal activity of AITC vapor in combination with cis-3-hexen-1-ol (c3H) vapor against Pd was examined.

[0134] Figures 25A-25E is a graph of the infection incidence in citrus. The antifungal activity of AITC in combination with c3H and essential oil vapor against Pd was examined.

[0135] Fig.26 is a graph of the normalized mean severity of lemon disks testing the antifungal activity of AITC, c3H, and cis-3-hexenyl acetate vapor against Pd.

[0136] Fig. 27 is a graph of the firmness of ripening characteristics of avocados after exposure to AITC vapor, after coating with an edible coating composition, after exposure to AITC vapor and coating with an edible coating composition, and a group of untreated avocados.

[0137] Fig.28 This is a graph showing the Shore hardness results of the four avocado test groups five days after Cg infection.

[0138] Fig.29 is a graph of Shore firmness of avocados, after 5 days of AITC vapor exposure and infection with Cg, the ripening characteristic, Shore firmness, was tested. DETAILED DESCRIPTION OF THE INVENTION

[0140] The present invention describes a slow-release gas phase treatment for mitigating latent infections, such as fungal infections, in agricultural products. Figure 1 is a flow chart showing steps in one embodiment of a method 100 for treating produce. At 102, a plurality of produce items are placed in an enclosure. At 104, an antimicrobial agent is provided to the enclosure. At 106, the plurality of produce items are contacted with the antimicrobial agent. The antimicrobial agent is gaseous and is selected to inactivate latent microorganisms in the plurality of produce items. At 108, after contacting the plurality of produce items with the antimicrobial agent for a period of time, the plurality of produce items are removed from the enclosure to produce a plurality of treated produce items.

[0141] Provided herein are methods for treating produce that include providing an antimicrobial agent to a housing containing a plurality of produce items; and contacting the plurality of produce items with the antimicrobial agent, wherein the antimicrobial agent is in gaseous form and is selected to inactivate latent microorganisms in the plurality of produce items.

[0142] New, more cost-effective methods are needed to prevent degradation by microorganisms, reduce the production of fungi / molds, maintain quality, and extend the life of agricultural products. The embodiments of the antimicrobial agent or the antimicrobial agent and edible coating composition described herein can provide one or more advantages. For example, in some embodiments, the antimicrobial agent or the antimicrobial agent and edible coating composition can protect the agricultural products from biological stress (bioticstressors), i.e., bacteria, viruses, fungi or pests. The antimicrobial agent or the antimicrobial agent and edible coating composition can also prevent water evaporation and / or the diffusion of oxygen, carbon dioxide and / or ethylene. The antimicrobial agent or the antimicrobial agent and edible coating composition can also help extend the shelf life of agricultural products (e.g., agricultural products after harvest) without refrigeration. The antimicrobial agent or the antimicrobial agent and edible coating composition can reduce the average mass loss rate of coated agricultural products. The antimicrobial agent or the antimicrobial agent and edible coating composition can also delay the maturity of the agricultural products. The antimicrobial agent or the antimicrobial agent and edible coating composition can also be of natural origin, so it is safe for human consumption. The methods of treating produce disclosed herein can extend the life of the produce and delay the ripening of the produce.

[0143] The enclosure may be selected to house two or more produce items. Examples of suitable enclosures include produce packaging containers (e.g., clamshells), produce storage containers (e.g., bins), produce transport containers (e.g., shipping containers or box trucks), and produce handling containers (e.g., chlorosis or ripening chambers).

[0144] Some embodiments include sealing the housing prior to providing the antimicrobial agent to the housing. Sealing the housing may include making the housing airtight, or limiting the diffusion rate of one or more selected gases into and out of the housing.

[0145] Providing the antimicrobial agent to the housing may include providing a gas flow comprising the antimicrobial agent to the housing. In one example, the gas flow comprising the antimicrobial agent is pumped into the housing.

[0146] The time length of contacting the multiple agricultural products items with the antimicrobial agent is within the range of 1 minute to 100 hours (e.g., 10 minutes to 1 hour). The time length depends on multiple factors, such as the type of agricultural products, the target pathogen and the sealing degree of the shell. Example includes the exposure time of allyl isothiocyanate (AITC) in a sealed box between 15 minutes and 96 hours. For the AITC concentration of 1000ppm, the exposure time of 15 minutes slows down the mold growth on the agricultural products, and the exposure time of 30 minutes or longer shows good results. It is also possible to use a lower antimicrobial concentration with a longer exposure time or a higher antimicrobial concentration with a shorter exposure time. For example, the time length of contacting the multiple agricultural products items with the antimicrobial agent is 15 minutes to 72 hours.

[0147] The concentration of the antimicrobial in the headspace of the shell is usually in the range of about 20 parts per million by volume (ppmv) to about 5000ppmv (for example, about 550ppmv to about 4800ppmv, or about 25ppmv to about 1250ppmv). Depending on the type of agricultural product and the exposure time in the shell, the antimicrobial concentration greater than about 2200ppmv may sometimes damage the agricultural product. When there is one of multiple synergistic effects, a lower antimicrobial concentration (for example, 25ppmv) may be more effective. For the AITC concentration in the range of 550ppmv to 4800ppmv, all concentrations have a certain inhibitory effect on mold growth, and the concentration exceeding 1000ppmv tends to be completely inhibitory. In some embodiments, the antimicrobial is provided to the shell in the headspace with a concentration of at least about 1200ppmv, at least about 1250ppmv, or about 1250ppmv to about 5000ppmv, or about 1250ppmv to about 4900ppmv.

[0148] Some embodiments include regulating the temperature or humidity inside the shell. In some embodiments, the temperature inside the shell is in the range of about 4°C to about 80°C (e.g., about 4°C to about 55°C), depending at least in part on the type of agricultural product to be treated and the antimicrobial agent used. In some embodiments, the relative humidity (RH) inside the shell is in the range of about 5% to about 99% (e.g., about 30% to about 98%). In some instances, the temperature is in the range of about 18°C ​​to about 24°C, and the relative humidity (RH) is in the range of about 40% to about 60%. Higher humidity and lower temperature generally reduce the concentration of passive volatile gases in the head space. Antimicrobial agents with lower volatility than AITC and DADS may reach effective concentrations in shells with temperatures above 23.33°C and / or relative humidity below 40%.

[0149] Exposure time and antimicrobial agent concentration, as well as temperature, relative humidity and other factors, can be selected to optimize antimicrobial performance for the produce being treated and the type of microorganisms being inactivated.

[0150] In some embodiments, an antimicrobial agent precursor is volatilized to produce the antimicrobial agent. The volatilization of the antimicrobial agent precursor may occur outside the shell or inside the shell.

[0151] Some embodiments include activating an antimicrobial precursor to produce the antimicrobial. Activating the antimicrobial precursor can include contacting the antimicrobial precursor with a catalyst selected to promote the conversion of the antimicrobial precursor into the antimicrobial. The catalyst can be a biocatalyst (e.g., an enzyme). In one example, the antimicrobial precursor comprises alliin, the enzyme comprises alliinase, and the antimicrobial comprises allicin. In another example, the antimicrobial comprises sinigrin, the enzyme comprises myrosinase, and the antimicrobial comprises allyl isothiocyanate.

[0152] Contacting the plurality of produce items with the antimicrobial agent can include circulating the antimicrobial agent in the housing (e.g., using pressurized air). Contacting the plurality of produce items with the antimicrobial agent can include penetrating an epidermal layer of each of the plurality of produce items with the antimicrobial agent. In some cases, the antimicrobial agent or a derivative thereof can be detected within a depth below the epidermal layer of the treated produce.

[0153] Non-limiting antimicrobial agents can include antibacterial agents, antifungal agents, antiviral agents or their combinations in any. The example of suitable antifungal agents includes allyl isothiocyanate, diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranilate, methyl cinnamate, γ-decanoic acid lactone, α-terpineol, linalool or its combinations in any. In some cases, the antifungal agent includes a derivative (for example, cis-3-hexenol or cis-3-hexene acetate) of 3-hexene. In some embodiments, the antimicrobial agent includes allyl isothiocyanate. The antifungal agent is selected to inactivate green mold or penicillium (e.g., Botrytis cinerea, Penicillium spp., Monilinia spp., Alternaria alternata, Rhizopus stolonifera, Trichothecium roseum, Fusarium spp., Colletotrichum spp., or any combination thereof). In some embodiments, the antimicrobial agent is selected to inactivate Colletotrichum spp. (e.g., Colletotrichum gloeosporioides, Cg).

[0154] In some embodiments, the antimicrobial agent comprises allyl isothiocyanate and one or more antimicrobial agents selected from the group consisting of diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranilate, methyl cinnamate, gamma-decalactone, alpha-terpineol, and linalool.

[0155] The plurality of agricultural products include fruits, vegetables, or combinations thereof. Suitable examples of fruits include berries (e.g., avocados, bananas, blueberries, cranberries, eggplants, tomatoes, grapes, persimmons), citrus fruits (e.g., oranges, lemons, limes, grapefruits, kumquats), gourds (e.g., pumpkins, cucumbers, watermelons), stone fruits (e.g., peaches, plums, cherries, olives), pome fruits (e.g., apples, pears), aggregate fruits (e.g., blackberries, raspberries), accessory fruits (e.g., strawberries), or any combination thereof. In some embodiments, the fruit is an avocado, strawberry, citrus, or lemon. In some embodiments, the fruit is an avocado.

[0156] Figure 22 is a schematic diagram of an embodiment of a system 200 configured to treat agricultural products with a vapor-phase antimicrobial agent to reduce latent infection. The system 200 includes a reactor 202 and a housing 204. The reactor 202 is configured to promote the conversion of an antimicrobial precursor into an antimicrobial agent. The reactor 202 includes a bed 206 configured to accommodate a granular medium coupled (e.g., covalently or non-covalently bound) to a catalyst, wherein the catalyst is selected to promote the conversion of the antimicrobial precursor to the antimicrobial agent. In some embodiments, the reactor 202 is a fixed bed reactor. The inlet 208 is near the first end of the bed 206 and is configured to guide the carrier gas from the first end of the bed to the second end of the bed. The additional inlet 210 is configured to guide the antimicrobial precursor to the bed 206; in some embodiments, the antimicrobial precursor is provided to the reactor 202 by a pump 212. The outlet 212 is near the second end of the bed 206 and is configured to allow a gas mixture containing the carrier gas and the antimicrobial agent to go out. The housing 204 is configured to be in fluid communication with the reactor 202 via the outlet 212. The housing is configured to receive a plurality of agricultural products and to contain a gas mixture in contact with the plurality of agricultural products for a period of time. In some embodiments, the housing 204 is a closed housing, such as a bio-fumigation chamber.

[0157] The biocide precursor in the vessel 214 is provided to the reactor 202 by pump 216, and the pump is connected with the reactor 202 fluids by extra inlet 210. In an example, pump 216 is a peristaltic pump. Extra inlet 210 can be arranged to provide the biocide precursor in a mode perpendicular to the length of bed 206. The catalyst coupled with the granular medium in bed 206 converts the biocide precursor into corresponding biocide. Carrier gas from source 218 is provided by inlet 208, and flows along the length of bed 206 (for example, from the first end of bed 206 to the second end of bed 206), volatilizes the biocide and forms a gaseous mixture comprising the carrier gas and biocide. In some embodiments, the carrier gas is the pressurized air supply provided to the reactor 202 as a pressurized gas or mechanical pressurized air supply.

[0158] The catalyst in reactor 202 can be a biocatalyst (e.g., an enzyme). In one example, the biocide precursor comprises alliin, the enzyme comprises alliinase, and the biocide comprises allicin. In another example, the biocide comprises sinigrin, the enzyme comprises sinigrinase, and the biocide comprises allyl isothiocyanate.

[0159] In some embodiments, the agricultural product processing method may also include coating the multiple agricultural product items with an edible coating composition. Coating the multiple agricultural product items with the edible coating composition may be before providing the antimicrobial agent to the shell, or after contacting the multiple agricultural product items with the antimicrobial agent. The edible coating composition may be applied to the surface of the agricultural product by means generally known in the art (e.g., dip coating). For example, a description of the method for coating agricultural products is provided, see, for example, U.S. Patent Publication No. 20190269144A1, which is incorporated herein by reference in its entirety. The edible coating composition may include a coating agent and a solvent, wherein the coating agent includes one or more saturated glycerides selected from monoglycerides and diglycerides; and one or more fatty acid salts.

[0160] In some embodiments, the edible coating composition includes one or more monoglycerides. In some embodiments, the coating agent includes one monoglyceride (e.g., 1-monoglyceride or 2-monoglyceride). In some embodiments, the coating agent includes two monoglycerides (e.g., two 1-monoglycerides, two 2-monoglycerides, or one 1-monoglyceride and one 2-monoglyceride). In some embodiments, the coating agent includes three monoglycerides.

[0161] In some embodiments, one or more of the monoglycerides have a carbon chain length of about C10 to about C22. In some embodiments, the monoglycerides have a carbon chain length of about C10 to about C22. In some embodiments, the carbon chain length of the monoglycerides includes one or more of the following, or is selected from the group consisting of: C10 monoglycerides, C12 monoglycerides, C14 monoglycerides, C16 monoglycerides, C18 monoglycerides, C20 monoglycerides, and C22 monoglycerides. In some embodiments, the monoglycerides are saturated monoglycerides. In some embodiments, the saturated monoglycerides are monolaurin, glyceryl monostearate, glyceryl monostearate, glyceryl monobehenate, glyceryl monobehenate, or glyceryl hydroxystearate. In some embodiments, the monoglyceride is glyceryl monostearate.

[0162] In some embodiments, the one or more glycerol monoesters are present in the coating agent in an amount ranging from about 40 wt% to about 99 wt%, based on the total weight of the coating agent. For example, the one or more glycerol monoesters are present in the coating agent in an amount ranging from about 50 wt% to about 98 wt%, from about 60 wt% to about 99 wt%, from about 70 wt% to about 98 wt%, from about 85 wt% to about 98 wt%, from about 90 wt% to about 98 wt%, from about 92 wt% to about 97 wt%, or from about 95 wt%, based on the total weight of the coating agent. In some embodiments, the one or more glycerol monoesters are present in the coating agent in an amount ranging from about 75 wt% to about 98 wt%, based on the total weight of the coating agent.

[0163] In some embodiments, the edible coating composition includes one or more fatty acid salts. In some embodiments, the coating agent includes a fatty acid salt. In some embodiments, the coating agent includes two fatty acid salts. In some embodiments, the coating agent includes three fatty acid salts. In some embodiments, the coating agent includes four or more fatty acid salts.

[0164] In some embodiments, at least one of the one or more fatty acid salts comprises a carbon chain length of about C10 to about C22. In some embodiments, each of the one or more fatty acid salts comprises a carbon chain length of about C10 to about C22. In some embodiments, at least one of the one or more fatty acid salts comprises a carbon chain length selected from: C10, C12, C14, C16, C18, C20 or C22. In some embodiments, each of the one or more fatty acid salts comprises a carbon chain length selected from: C10, C12, C14, C16, C18, C20 or C22. In some embodiments, the one or more fatty acid salts are C14 fatty acid salts, C16 fatty acid salts, C18 fatty acid salts, or combinations thereof. In some embodiments, the one or more fatty acid salts are C16 fatty acid salts and C18 fatty acid salts. In some embodiments, one or more of the fatty acid salts are saturated. In some embodiments, one or more of the fatty acid salts are unsaturated.

[0165] In some embodiments, one or more of the fatty acid salts is a salt of lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, palmitoleic acid, caprylic acid, capric acid, cerotic acid, oleic acid, linoleic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, myristoleic acid, sapienic acid, elaidic acid, vaccenic acid, linoelaidic acid, alpha-linolenic acid, erucic acid, docosahexaenoic acid, or a combination thereof.

[0166] In some embodiments, the fatty acid salt comprises one or more of the following, or is selected from the group consisting of sodium laurate, sodium myristate, sodium palmitate, sodium stearate, arachidate, sodium behenate, sodium lignocerate, sodium arachidonic acid, eicosapentaenoic acid sodium salt, docosahexaenoic acid sodium salt, sodium myristate, sapienate, elaidate, linoleic acid sodium salt, linoleic acid sodium salt, sodium erucate and docosahexaenoic acid sodium salt.

[0167] In some embodiments, the one or more fatty acid salts are present in the coating agent in an amount in the range of about 1 wt % to about 20 wt %, based on the total weight of the coating agent. For example, the one or more fatty acid salts are present in the coating agent in an amount in the range of about 1 wt % to about 10 wt %, about 1 wt % to about 8 wt %, 1 wt % to about 5 wt %, or about 1 wt % to about 3 wt %, based on the total weight of the coating agent. In some embodiments, the one or more fatty acid salts are present in the coating agent in an amount in the range of about 1 wt % to about 5 wt %, based on the total weight of the coating agent.

[0168] In some embodiments, the coating agent comprises one or more glycerol monoesters in an amount ranging from about 90 wt% to about 98 wt%, and one or more fatty acid salts in an amount ranging from about 1 wt% to about 8 wt%. In some embodiments, the coating agent comprises glycerol monostearate in an amount ranging from about 90 wt% to about 98 wt%, and sodium stearate in an amount ranging from about 1 wt% to about 8 wt%.

[0169] In some embodiments, the coating agent further comprises an antimicrobial agent as disclosed herein. In some embodiments, the coating agent further comprises one or more antimicrobial agents, wherein the one or more antimicrobial agents are selected from allyl isothiocyanate, diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranilate, methyl cinnamate, γ-decanoic acid, α-terpineol, linalool, cis-3-hexenol or cis-3-hexene acetate, sodium benzoate, potassium sorbate, chalcone, fludioxonil, 2-hydroxychalcone, 4-hydroxychalcone, Chalcone, 4'-hydroxychalcone, 2,2'-dihydroxychalcone, 2,4'-dihydroxychalcone, 2',4-dihydroxychalcone, 2',4'-dihydroxychalcone, 2',4,4'-trihydroxychalcone, 2',4,4'-trihydroxychalcone intermediate, violastyrene, obtusaquinone, apiole, piperine, celastrol, arthonoic acid acid, leoidin, antimycin A, antimycin A1, refractory lichenic acid, ethyl orsellinate, methylorsellinate, mycophenolic acid, ethyl dichloroorsellinate, angoratin, isocarpone, eupatoriochromene, peppermint, lichenic acid, aloesin, formononetin, acetovanillone, isoorsentin, deoxysedanone B 7,4'-dimethyl ether, chrysanthemum yellow ether, bergamot, garcinia acid, 2-hydroxyxanthone, isopimpinellin, peppermint, acetyl hymetochrome, nobiletin, hymechrome, methoxsalen, 4-methylesculetin, tangerin, kaelin, flavonoids, 3,4',5,6,7-pentamethoxyflavone, deguelin (-), limonene, deoxyserotonin B trimethyl ether, deoxyserotonin B 7,3'-dimethyl ether, 2',4'-dihydroxy-4-methoxychalcone, daunorubicin hydrochloride, plumbagin, menadione, musciquinone, levomenthol, methyl trimethoxycinnamate, chavicol, cinnamylphenol, benzoate, napthoquinone, benzophenone, acetophenone, benzophenone, phenylacetophenone, chitosan, salicylic acid and sodium salicylate.

[0170] In some embodiments, the coating agent can be dissolved, mixed, dispersed or suspended in a solvent to form a composition (e.g., a solution, a suspension or a colloid). Examples of usable solvents include water, methanol, ethanol, isopropanol, butanol, acetone, ethyl acetate, chloroform, acetonitrile, tetrahydrofuran, diethyl ether, methyl tert-butyl ether or a combination thereof. For example, the solvent is water. For example, the solvent is ethanol.

[0171] In some embodiments, the coating agent is present in an edible coating composition (e.g., solution, suspension, or colloid) at a concentration of about 1 g / L to 200 g / L. For example, about 1 to 150 g / L, 1 to 100 g / L, 1 to 75 g / L, 1 to 150 g / L, 1 to 50 g / L, 1 to 25 g / L, 10 to 200 g / L, 10 to 150 g / L, 10 to 100 g / L, 10 to 75 g / L, 10 to 50 g / L, 10 to 45 g / L, 10 to 40 g / L, 25 to 75 g / L, 35 to 65 g / L, 40 to 60 g / L, or 45 to 55 g / L.

[0172] In some embodiments, any edible coating composition described herein may further comprise one or more additives. In some embodiments, the additive comprises one or more of the following, or is selected from the group consisting of: preservatives, stabilizers, buffers, vitamins, minerals, pH regulators, salts, pigments, flavors, enzymes, catalysts, antioxidants, antifungal agents, antimicrobial agents, or combinations thereof.

[0173] In some embodiments, the edible coating composition comprises a coating agent comprising one or more glycerol monoesters in an amount ranging from about 90 wt% to about 98 wt%, and one or more fatty acid salts in an amount ranging from about 1 wt% to about 8 wt%, and a solvent, wherein the concentration of the coating agent in the edible coating composition is about 25 g / L to 75 g / L. In some embodiments, the edible coating composition comprises a coating agent comprising glycerol monostearate in an amount ranging from about 90 wt% to about 98 wt%, and sodium stearate in an amount ranging from about 1 wt% to about 8 wt%, and a solvent comprising water, wherein the concentration of the coating agent in the edible coating composition is about 25 g / L to 75 g / L.

[0174] Also provided herein are a plurality of agricultural products items processed by the agricultural products processing method disclosed herein. When the plurality of agricultural products items are processed by the agricultural products processing method disclosed herein, the plurality of agricultural products items may have an increase in the mass loss factor. For example, the mass loss factor of a plurality of agricultural products items not processed by the agricultural products processing method disclosed herein is 1, and the mass loss factor of a plurality of agricultural products items processed by the agricultural products processing method disclosed herein is at least 1.1, or at least 1.5, or at least 1.7, or at least 1.8, or at least 2, or at least 2.2, or at least 2.5, or in the range of 1.1 to 2.8, or in the range of 2 to 2.6. For example, the mass loss can be measured by determining the weight difference of the agricultural products after the agricultural products are processed by the agricultural products processing method disclosed herein and after a period of time. In some embodiments, the mass loss is measured after 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, days, 8 days, 9 days and / or 10 days or any combination thereof. In some embodiments, the mass loss is measured after 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, or any combination thereof. Advantageously, the plurality of produce items treated by the produce treatment methods disclosed herein have a higher mass loss factor than the untreated plurality of produce items.

[0175] The plurality of produce items can be at least 5 (e.g., at least 5, 10, 15, 20, 25, 30, 40, 50) avocados (e.g., California avocados, Mexican avocados, etc., and the avocados can be #84, #70, #60, #48, #40, #36, #32, #28 avocados, etc.). For example, the plurality of produce items are processed California #60 avocados (e.g., 30 California #60 avocados), wherein the processed California #60 avocados are California #60 avocados that have been processed by the produce processing methods disclosed herein. In some embodiments, wherein the plurality of produce items are 30 treated California #60 avocados, and 5 days after the 30 treated California #60 avocados are treated, the 30 treated California #60 avocados have an average Shore value that is at least 10 Shores, at least 15 Shores, at least 20 Shores, or at least 25 Shores higher than the average Shore value of 30 untreated California #60 avocados, and wherein on day 0, the 30 treated California #60 avocados have an average Shore value that is ±10 Shores of the 30 untreated California #60 avocados. As used herein, the term "day 0" refers to the day that the plurality of produce items are treated, but before the plurality of produce items are actually treated.

[0176] In some embodiments, after the agricultural product is treated by the agricultural product treatment methods disclosed herein, an amount of green mold or penicillium in the agricultural product is inactivated. For example, the treatment of the agricultural product by the agricultural product treatment methods disclosed herein can be used to prevent the growth of green mold or penicillium (e.g., Botrytis cinerea, Penicillium spp., Monilinia spp., Alternaria alternata, Rhizopus stolonifera, Trichothecium roseum, Fusarium spp., Colletotrichum spp., or any combination thereof).

[0177] In some embodiments, after the agricultural product is treated by the agricultural product treatment method disclosed herein, the respiration rate of the agricultural product can be reduced. For example, the treatment of agricultural products by the agricultural product treatment method disclosed herein can be used to prevent or limit the diffusion of gases (such as ethylene, CO2 and O2, etc.), thereby slowing down maturity and / or aging. In some embodiments, after the agricultural product is treated by the agricultural product treatment method disclosed herein, the rate at which the agricultural product produces CO2 is reduced.

[0178] Also provided herein are a plurality of agricultural product items coated with the edible coating composition, wherein the edible coating composition comprises a coating agent comprising: one or more monoglycerides; and one or more fatty acid salts; a solvent; and allyl isothiocyanate. The coating agent may be as described above, but the edible coating composition further comprises allyl isothiocyanate. In some embodiments, the allyl isothiocyanate is present in the edible coating composition in an amount of about 0.001% v / v to about 5% v / v. For example, the allyl isothiocyanate is present in the edible coating composition in an amount of about 0.01% v / v to about 2.5% v / v, about 0.1% v / v to about 1.5% v / v, or about 0.5% v / v to about 1.5% v / v.

[0179] In some embodiments, after applying the edible coating composition comprising allyl isothiocyanate, a certain amount of green mold or penicillium of the agricultural product is inactivated. For example, treatment with an edible coating composition comprising allyl isothiocyanate disclosed herein can be used to prevent the growth of green mold or penicillium (e.g., Botrytis cinerea, Penicillium, Sclerotinia, Alternaria interspersata, Rhizopus glabrata, Trichothecene rosea, Fusarium, Trichothecene, or any combination thereof).

[0180] In some embodiments, after the edible coating composition is applied, the respiration rate of the agricultural product can be reduced. For example, the application of any edible coating composition described herein can be used to prevent or limit the diffusion of gases (such as ethylene, CO2 and O2, etc.), thereby slowing down maturation and / or aging. In some embodiments, after the edible coating composition or emulsion is applied, the rate at which the agricultural product produces CO2 is reduced.

[0181] The above description and the following examples describe in detail certain specific embodiments of the present invention, and describe the best mode of implementation contemplated by the inventors. However, it will be appreciated that no matter how detailed the above description appears in text, the present invention can be implemented in a variety of ways, and the present invention should be interpreted according to the appended claims and their equivalents.

[0182] Although the disclosed teachings have been described with reference to a variety of applications, methods, compounds, compositions and materials, it should be understood that they can be subjected to a variety of changes and modifications without departing from the teachings herein. The following examples are provided to better illustrate the disclosed teachings and are not intended to limit the scope of the teachings presented herein. Although the current teachings have been described according to these exemplary embodiments, it will be readily appreciated by those skilled in the art that many variations and modifications of these exemplary embodiments are possible without the need for excessive experimentation. All such variations and modifications are within the scope of the teachings of the present invention. Example

[0183] Example 1. The antifungal activity of diallyl disulfide (DADS) vapor against Penicillium digitatum (Pd) was tested in citrus. All samples were wounded with a screw, and 1000 Pd spores suspended in 10 μL of water were distributed into each wound for infection. The test groups consisted of the following citrus: untreated; immersed in 1 g / L imazalil solution; and exposed to 3 mL DADS vapor for 72 hours in a Petri dish. Figure 3 The infection quantification shown indicates that exposure to DADS vapor reduced the incidence of Pd mold by 95.8% compared to untreated samples and by 19.1% compared to samples treated with imazalil. There was no visible degradation or discoloration of citrus exposed to DADS vapor. Figure 3 It was shown that passive volatilization of DADS in a closed, unsealed bin significantly prevented Pd mold growth in citrus.

[0184] Example 2. The antifungal activity of allyl isothiocyanate (AITC) and DADS vapor against Pd and Penicillium italicum (Pi) was tested in citrus. All samples were wounded by piercing with a screw. Citrus was infected by distributing 1000 Pd spores or 1000 Pi spores suspended in 10 μL into the wound. The test group consisted of the following citrus: uninfected and untreated; uninfected and treated by immersion in a 1 g / L imazalil solution; infected with Pd and untreated; infected with Pd and exposed to AITC; infected with Pd and exposed to DADS; infected with Pi and untreated; infected with Pi and exposed to AITC; and infected with Pi and exposed to DADS. Exposure to AITC or DADS consisted of exposing the citrus to 3 mL of the vapor of the corresponding compound distributed in a Petri dish for 72 hours. Figure 4 Quantification of the test results is shown. AITC vapor substantially prevented both Pd and Pi infection. Citrus treated with AITC appeared visibly darker than untreated fruit. DADS vapor reduced the incidence of Pd mold by 85% and Pi mold by 30% compared to the corresponding untreated / infected test groups. No differences in appearance were observed for fruit treated with DADS compared to untreated samples. Both AITC and DADS were effective in preventing fungal growth. AITC had approximately 100% inhibition with some changes in visible appearance.

[0185] Example 3. The antifungal activity of decreasing doses of AITC vapor against Pd was tested in citrus. All samples were wounded by puncturing with a screw. Citrus was infected by dispensing 1000 Pd spores suspended in 10 μL of water into each wound. The test groups consisted of the following citrus: uninfected and untreated; infected with Pd and untreated; and exposed to 500 μL AITC vapor for 72 hours in a Petri dish. Figure 5 Quantification of infection of the test group is shown. The uninfected and untreated group showed significant infection in initially uninfected fruit. AITC doses of 1 / 6 of the dose used in Example 2 substantially prevented Pd infection. Spotting and darkening of the fruit skin were observed in some samples of the AITC treated fruit.

[0186] Example 4. A range of AITC vapor concentrations were tested for antifungal activity against Pd in ​​citrus. All citrus were wounded with screws, and 1000 Pd spores suspended in 10 μL of water were distributed into each wound for infection. The fruit was then placed in a sealed container, a range of AITC volumes were arranged on a Whatman filter paper disc, the filter disc was suspended in front of a battery-powered fan, and the container was sealed for 18 hours. The citrus was then placed at ambient temperature and high humidity for 6 days. The test groups were infected and untreated citrus, as well as infected and treated with 62.5 μL, 125 μL, 250 μL, and 500 μL on the filter paper disc. The infection in the 62.5 μL treatment group was lighter than that in the control group. Figure 6 is a bar graph showing the morbidity (percent infection) of the test groups. The 62.5 μL dose of AITC reduced the severity of infection relative to the untreated group. The 125 μL, 250 μL and 500 μL doses of AITC substantially prevented mold growth on the treated citrus. The 250 μL and 500 μL doses resulted in darkening of the peel.

[0187] Example 5. Antifungal activity over a range of AITC vapor exposure times was tested to determine the effect against Pd in ​​citrus. All samples were wounded with a screw. Citrus was infected by dispensing 1000 Pd spores suspended in 10 μL of water in the wound. The fruit was then placed in a sealed container, 125 μL of AITC was dispensed onto a Whatman filter paper disc, the filter disc was suspended in front of a battery-powered fan, and the container was sealed for 15 minutes to 2 hours. The test group was then placed at ambient temperature, high humidity, and evaluated for Pd mold after 6 days. The test group consisted of the following citrus: uninfected and untreated; infected and untreated; infected and treated with 0.5 g / L imazalil; infected, treated with 125 μL AITC on filter paper, and exposed to vapor for 15 minutes, 30 minutes, 1 hour, and 2 hours. Figure 7 Quantified infection results are shown. A 15 minute exposure had little effect on Pd mold, a 30 minute exposure greatly reduced the severity of Pd mold infection, a 1 hour exposure greatly reduced the severity and incidence of Pd mold, and a 2 hour exposure virtually eliminated mold. Latent fungal infection other than Pd confounded the results of the imazalil control and the 1 hour AITC treatment. For the 2 hour treatment, spotting and darkening of the peel was observed in 62.5% of the fruit. Exposure to 730 ppm concentration of AITC vapor for 30 minutes appears to virtually prevent mold infection or reduce the severity of infection. Longer exposure times increase the antifungal effect. As shown in the 1 hour exposure samples ( Figure 7) observed that latent pathogens were substantially resistant to AITC vapor at this concentration and exposure time. Longer exposure times were effective in substantially eliminating these pathogens.

[0188] Example 6. The antifungal activity of AITC vapor was measured using a range of incubation times after infection before treatment to detect the treatment window against Pd in ​​citrus. All samples were wounded with screw punctures and 1000 Pd spores suspended in 10 μL of water were distributed in the wounds for infection. The citrus was then incubated for 0 to 4 days before treatment. For treatment, the citrus was placed in a sealed container, 125 μL AITC was distributed on a Whatman filter paper disc, the filter disc was hung in front of a battery-powered fan, and the container was sealed for 30 minutes. The test group was then placed at ambient temperature, high humidity, and Pd mold was evaluated after 6 days. The test group consisted of the following citrus: untreated; treated with 1 g / L of imazalil; and incubated for 0 hours, 24 hours, 48 ​​hours, 72 hours, and 96 hours before AITC treatment. Figure 8 The quantified infection results are shown. The fruit quality was poor, with significant latent infection rates of multiple fungal pathogens (including Penicillium), and 83.3% were infected in the imazalil treated group. For those treated with AITC immediately after infection, 58.3% were infected. For those treated 24 hours later, 8.3% were infected. For those treated 48 hours later, 25% were infected. For those treated 72 hours later, 16.7% were infected. For those treated 96 hours later, 81.6% were infected. Treatment 24 hours after infection resulted in the lowest percentage of Penicillium infection in the test group. Treatment immediately after infection resulted in the worst fruit quality in the test treatments. It is not clear whether this is due to the treatment regimen or the initial fruit quality. Intermediate treatment time points appear to be effective against infection.

[0189] Example 7. The antifungal activity of AITC and DADS vapor against Colletotrichum gloeosporioides (Cg) was tested in avocado. Wounds were created in all test blocks by removing the stems and infected by dispensing 1000 Cg of spores suspended in 10 μL of water into the wounds. The test groups consisted of the following avocados: untreated and uninfected; untreated and infected; treated by immersion in a 0.1 g / L prochloraz solution; exposed to 3 mL of AITC vapor arranged in a Petri dish; and exposed to 3 mL of DADS vapor arranged in a Petri dish. Fig. 9Infection results were quantified. AITC vapor reduced the incidence of Cg mold by 97% compared to the untreated test group and by 70% compared to the prochlorperazine-treated test group. DADS vapor reduced the incidence of Cg mold by 60% compared to the untreated test group.

[0190] Example 8. The antifungal activity of Cg over a range of DADS vapor exposure times was tested in avocado. Wounds were created in all test plots by removing the stem and infected by dispensing 1000 Cg spores suspended in 10 μL of water into the wound. The test groups consisted of the following avocados: infected and untreated; and exposed to 3 mL DADS vapor for 24 hours, 48 ​​hours, and 72 hours in a Petri dish. Fig.10 The infection results were quantified. Test groups exposed to DADS vapor at all exposure times had reduced incidence of Cg mold compared to the untreated test group. 24-hour DADS exposure reduced incidence of Cg mold by 8%. 48-hour DADS exposure reduced incidence of Cg mold by 25%. 72-hour DADS exposure reduced incidence of Cg mold by 57%.

[0191] Example 9. The antifungal activity of Cg over a range of AITC and carvacrol vapor exposure times was tested in avocado. To measure the effect of the various treatments on ripening, Shore hardness was measured after 7 days of incubation at ambient temperature and high humidity. All samples were wounded by removing the stem and infected by dispensing 1000 Cg spores suspended in 10 μL of water into the wound. The test groups consisted of avocados that were infected and left untreated on the benchtop; infected and left untreated under the fume hood; exposed to 3 mL AITC vapor in a Petri dish for 24 hours, 48 ​​hours and 72 hours; and exposed to 3 mL carvacrol vapor in a Petri dish for 72 hours. Fig.11 The infection outcomes were quantified. Fig.12 The Shore hardness results for the test groups are shown. All exposure times to AITC vapor reduced the incidence of Cg mold compared to the untreated samples. 24 hours of AITC exposure reduced the incidence of Cg mold by 90%. 48 hours of AITC exposure reduced the incidence of Cg mold by 97%. 72 hours of AITC exposure reduced the incidence of Cg mold by 83%. Exposure to carvacrol vapor did not reduce the incidence of Cg mold. Fig.12 All AITC treatments showed significantly higher Shores than untreated avocados (mean increase of 20.3 Shores; P value < 0.0001).

[0192] Example 10. A range of AITC vapor exposure times was tested in avocado for the antifungal activity of Cg. To measure the effect of various treatments on ripening, Shore hardness was measured after 6 days of incubation at ambient temperature and high humidity. All samples were wounded by removing the stem and infected by dispensing 1000 Cg spores suspended in 10 μL of water into each wound. The test groups consisted of avocados that were infected and left untreated on the benchtop; infected and left untreated in the fume hood; and exposed to 3 mL AITC vapor in a Petri dish for 24 hours, 48 ​​hours, and 72 hours. Fig.13 The infection outcomes were quantified. Fig.14 The Shore hardness results for the test groups are shown. All exposure times to AITC vapor reduced the incidence of Cg mold compared to the untreated samples. 24 hours of AITC exposure reduced the incidence of Cg mold by 81.5%. 48 hours of AITC exposure reduced the incidence of Cg mold by 93.5%. 72 hours of AITC exposure reduced the incidence of Cg mold by 86.5%. All AITC treatments had significantly higher Shores than infected avocados (average increase of 15.4 Shores; P value <0.005).

[0193] Example 11. The ripening characteristics of avocados exposed to AITC vapor were analyzed. The treatment groups consisted of the following avocados: exposed to 125 μL AITC vapor for 30 minutes and 18 hours in Petri dishes; exposed to 500 μL AITC vapor for 30 minutes and 18 hours; untreated and stored in a bin for 18 hours; and treated with a coating agent. Two groups of the above test groups were studied. One group was treated with coated fruit when cold, i.e. fruit treated with a coating agent (95 wt% glycerol monostearate and 5 wt% sodium stearate), and the other group was treated when warm (one day after being removed from refrigeration). Respiration and hardness (using a durometer) were measured daily. At the beginning of the test, the avocados were placed in the refrigerator. One day later, the cold-treated avocados were treated with AITC directly after being removed from the refrigerator. The coated group was treated at this time. The warm-treated avocados were allowed to warm to room temperature. On day 2, measurements were started on the cold treated avocados, and AITC treatment was started on the warm treated samples. On day 3, measurements were started on the warm treated group. The test groups consisted of the following avocados: untreated; exposed to 500 μL AITC vapor for 18 hours; and exposed to 125 μL AITC vapor for 30 minutes. At the end of the test, the avocados were cut and each half was imaged in full plane at less than 40 Shore. Fig.15is an image of an avocado half exposed to 500 μL AITC vapor. The peels of these high-level treatment test groups were thicker than indicated by the firmness meter and the flesh was more mature than the low-level treatment groups. Fig.16 A schematic diagram of the enzyme pathways hypothesized to be associated with thickening of the peel is shown. Without being bound by theory, upregulation of defense response pathways may trigger the production of defense compounds and antioxidants (e.g., caffeic acid, ferulic acid, sinapinic acid, and anthocyanins), potentially including lignin. This upregulation of lignin may result in thickening of the peel of avocados, which would give the perception of a firmer fruit, while the flesh inside has ripened and softened normally. These results indicate that exposure of the test group to AITC vapor at a level of 500 μL resulted in darkening and thickening of the avocado peel, as well as prolonged perception of firmness. The maintenance of firmness was not supported by changes in respiration rate, which would indicate any changes in fruit ripening or aging.

[0194] Example 12. The antifungal activity of AITC and DADS vapor against Botrytis cinerea (Bc) was tested in grapes. All samples were wounded by making a 3 mm deep hole in the skin. The samples were infected by dispensing 1000 Bc spores suspended in 10 μL of water in the wound. The test groups consisted of the following grapes: uninfected and untreated; infected and untreated; exposed to 3 mL AITC vapor for 72 hours in a Petri dish; and exposed to 3 mL DADS vapor for 72 hours in a Petri dish. Fig.17 The infection results were quantified. Exposure to AITC and DADS vapors virtually eliminated Bc mold. Grapes exposed to DADS vapor appeared unchanged after incubation, with no visible differences compared to untreated and uninfected grapes. Grapes exposed to AITC vapor showed darkening of the fruit. Both AITC vapor and DADS vapor were effective in controlling Bc mold. Treatment with AITC could potentially result in changes in grape color or flavor.

[0195] Example 13. The antifungal activity of AITC vapor against Bc was detected in strawberries. All samples were wounded by making 3 mm deep holes in the skin. The samples were infected by distributing 1000 Bc spores suspended in 10 μL of water in the wound. Then, the samples were incubated, imaged and evaluated for mold at ambient temperature over the next 3 days. The test group consisted of the following strawberries: uninfected and untreated; infected and untreated; and exposed to 125 μL AITC vapor distributed on Whatman filter paper for 30 minutes. After 24 hours, no mold was seen on these samples. After 48 hours, 5 / 8 of the uninfected samples showed visible mold, 5 / 8 of the infected samples showed visible mold, and 1 / 8 of the samples treated with AITC vapor showed visible mold. After 72 hours, 7 / 8 of the uninfected samples showed visible mold, 7 / 8 of the infected samples showed visible mold, and 1 / 4 of the samples treated with AITC vapor showed visible mold. Fig.18 Quantification of the infection results is shown. AITC vapor was effective against Bc mold in strawberries stored under ambient conditions. The infection rate in initially uninfected samples suggests that the steps involved in initial infection by distribution of Bc spores to the wound may not be necessary to promote infection in the test group.

[0196] Example 14. A range of AITC vapor concentrations were tested in strawberries for antifungal activity against Bc. Samples were treated by exposure to a range of volumes of AITC vapor arranged on filter paper discs for 30 minutes, then incubated at ambient temperature, imaged, and evaluated for mold over the next 6 days. The test groups consisted of untreated strawberries and strawberries exposed to vapors of 62.5 μL, 125 μL, 250 μL, and 500 μL AITC. Fig.19 Quantification of the infection results is shown. On day 6, 83% of the untreated control strawberries were infected, 100% of the 62.5 μL AITC dosed strawberries were infected, 13% of the 125 μL AITC dosed strawberries were infected, and 0% of the 250 and 500 μL AITC dosed strawberries were infected. For strawberries incubated at room temperature, all AITC doses above 62.5 μL were effective in preventing mold, and doses above 125 μL virtually eliminated mold.

[0197] Example 15. The antifungal activity of AITC vapor against Bc was tested in strawberries under different exposure conditions. The test groups consisted of untreated strawberries and strawberries treated by exposure to 125 μL of AITC vapor placed on a filter paper disk for 30 minutes. The test groups were then incubated in an open container or a closed container at 4°C, imaged, and evaluated for mold over the next 14 days. Fig. 20Quantification of the infection results is shown. All samples were free of mold within 7 days. On day 12, 16.7% of the open controls were infected, 29.2% of the closed container controls were infected, and 0% of the AITC treated samples were infected. On day 14, 33.3% of the open controls were infected, 41.7% of the closed container controls were infected, 0% of the open AITC treated samples were infected, and 4.2% of the closed container AITC treated samples were infected. For strawberries incubated at 4°C, AITC treatment was effective in reducing the incidence of mold over a two-week period. 1 / 3 of the open untreated samples were infected, while none of the open AITC treated samples were infected. Over 40% of the closed container controls were infected. One strawberry was infected in the closed container AITC treated test group.

[0198] Example 16. The antifungal activity of AITC vapor against Bc was tested in strawberries treated with a coating agent (95 wt% glyceryl monostearate and 5 wt% sodium stearate) and exposed to vapor in a clamshell. The test groups consisted of the following strawberries: untreated; immersed in a 40 g / L solution of the coating agent; exposed to 125 μL AITC vapor disposed on a filter paper disc for 30 minutes; and immersed in a 40 g / L solution of the coating agent and exposed to 125 μL AITC vapor disposed on a filter paper disc for 30 minutes. Over the next 4 days, the samples were treated, incubated at ambient temperature, imaged, and evaluated for mold. Fig.21 Quantification of the infection results is shown. On the 3rd day, 52.8% of the control group was infected, 42.1% of the coating group was infected, 26.3% of the AITC group was infected, and 9.3% of the coating+AITC group was infected. On the 4th day, 88.9% of the control group was infected, 76.3% of the coating group was infected, 71% of the AITC group was infected, and 34.9% of the coating+AITC group was infected. Exposing the strawberries to AITC vapor in a flip lid was less effective than exposing the strawberries in a more dispersed manner as described in the previous examples. Bowl immersion in the coating solution also poses a challenge to drying around the calyx. Both AITC-treated strawberries and coating+AITC-treated strawberries had significantly lower mold incidence rates than the untreated group.

[0199] Example 17. The antifungal activity of AITC vapor against Bc was tested in strawberries treated with a coating agent and hung. The test groups consisted of the following strawberries: untreated; hung and immersed in a 50 g / L solution of the coating agent; exposed to a vapor of 125 μL AITC placed on a filter paper disc for 30 minutes; hung and immersed in a 50 g / L coating agent solution and exposed to a vapor of 125 μL AITC placed on a filter paper disc for 30 minutes. Over the next 5 days, the samples were hung, treated, incubated at ambient temperature, imaged, and evaluated for mold. Fig. 22 is a graph of the mass loss factor (MLF) of the test group. The mass loss rate % (%MLR) is the percentage of mass loss of the produce group relative to itself per day. The mass loss factor (MLF) is the ratio of the untreated %MLR to the treated (e.g., coating agent-applied) %MLR. Fig.23 is a graph of the percentage of mold measured for the test groups. Hanging strawberries for treatment and incubation reduced accumulation under the calyx and generally improved drying relative to unhanged conditions. Fruits treated with the coating and the coating + AITC had significantly higher MLFs compared to the control. By day 5, AITC-treated strawberries and the coating + AITC-treated strawberries had significantly less mold than the untreated test group, with incidences reduced by more than 80% and 60%, respectively. Firmness measurements showed that all treated strawberries had improved firmness relative to the untreated test group. Time-lapse videos showed that both coating-treated test groups had better color retention and less shrinkage compared to the other test groups. Only the coating-treated test group had some signs of mold on day 5. No mold was observed in the coating + AITC-treated test group.

[0200] Example 18. The antifungal activity of AITC in combination with cis-3-hexen-1-ol (c3H) vapor against Pd was tested in citrus. All samples were punctured with a screw to create a wound. The samples were infected by dispensing 1000 Pd spores suspended in 10 μL of water into the wound. Exposure to vapor was accomplished by placing the citrus in a sealed container, dispensing AITC or AITC+c3H onto a Whatman filter paper disc, hanging the filter paper disc in front of a battery-powered fan, closing the container and incubating the citrus for 30 minutes. After vapor exposure, the citrus was placed at ambient temperature, high humidity, and evaluated for Pd mold on days 4-7. The test groups consisted of the following citrus: uninfected and untreated; infected and untreated; infected and treated by immersion in a 0.5 g / mL imazalil solution; exposed to 125 μL AITC vapor for 15 minutes to 30 minutes; exposed to 62.5 μL AITC and 125 μL c3H vapor for 30 minutes. Images of the untreated / infected, AITC-treated, and AITC+c3H test groups were taken on day 5. Images of the untreated / infected, AITC-treated, and AITC+c3H test groups were taken on day 6. Images of the untreated / infected, AITC-treated, and AITC+c3H test groups were taken on day 7. Fig.24 Quantification of infection results is shown. A 62.5 μL dose of AITC (50% of the standard dose) applied together with c3H resulted in a lower percentage of infection than a full strength dose of AITC alone. AITC+c3H reduced the incidence and severity of infection relative to the other treatments.

[0201] Example 19. The antifungal activity of AITC in combination with c3H and essential oil vapor against Pd was detected in citrus. All samples were punctured with a screw to create a wound, and 1000 Pd spores suspended in 10 μL of water were distributed in the wound for infection. Exposure to vapor was completed by the following steps: placing the citrus in a closed container, distributing the compound on a Whatman filter paper disc, hanging the filter paper disc in front of a battery-powered fan, closing the container and incubating the citrus for 30 minutes. After vapor exposure, the citrus was placed at ambient temperature and high humidity, and Pd mold was evaluated after 6 days. The test group not exposed to vapor consisted of untreated and treated citrus by immersion in a 1.0 g / mL imazalil solution. The remaining test groups were exposed to 62.5 μL of AITC vapor and / or 125 μL of all other compounds for 30 minutes. The compounds detected alone or in combination were AITC, c3H, cinnamaldehyde, eugenol and limonene. Figures 44A-44E show quantification of infection results. AITC vapor was applied at 50% of the minimum inhibitory concentration to detect synergy with any other tested essential oil. At this lower dose, AITC tended to reduce the severity of infection, but could not eliminate the incidence of infection, leaving room for improvement. Of the 15 test treatments, 8 performed better than the untreated sample. These treatments were: AITC+c3H; AITC+eugenol; AITC+limonene; cinnamaldehyde+c3H; cinnamaldehyde+eugenol; eugenol; eugenol+c3H; and eugenol+cinnamaldehyde.

[0202] Example 20. The antifungal activity of AITC, c3H, and cis-3-hexenyl acetate against Pd was tested in lemons. The test groups consisted of the following lemon disks: untreated; exposed to AITC vapor; exposed to c3H vapor; and exposed to cis-3-hexenyl acetate vapor. The results of infection severity in the samples were Fig.26 is quantified.

[0203] Example 21. The antifungal activity of avocados exposed to AITC vapor and prochloraz was tested against Cg in avocados. To measure the effect of various treatments on ripening, Shore hardness was measured after 5 days of incubation at ambient temperature (about 20°C to about 25°C) and high humidity (about 88% relative humidity to about 93% relative humidity). All samples were wounded by removing the stem and infected by dispensing 1000 Cg spores suspended in 10 μL of water into the wound. Exposure to vapor was accomplished by placing the avocados in a sealed container, dispensing AITC onto a Whatman filter paper disc, hanging the filter paper disc in front of a battery-powered fan, closing the container and incubating the avocados for 30 minutes. After vapor exposure, the avocados were placed at ambient temperature, high humidity, and evaluated for Cg mold on the 5th day. The test groups consisted of the following avocados: exposed to 125 μL of AITC vapor placed on a filter paper disk for 30 minutes; immersed in a 0.1 g / L prochloraz solution and placed on a rack in front of a fan to dry for 30 minutes; infected and untreated; and uninfected and untreated. Fig.28 The Shore hardness results for the test group are shown. To prepare 0.1 g / L of Prochloraz, 10 mg of Prochloraz was mixed with 10 mL of ethanol and 90 mL of water. AITC vapor reduced the incidence of Cg mold compared to the untreated sample and the Prochloraz-treated sample. 24-hour AITC exposure reduced the incidence of Cg mold by 90%. Fig.28 The results showed that the AITC treatment had significantly higher Shore values ​​than the untreated avocados, more than 30 Shore values. The average Shore value of the untreated avocados was 28.8 (standard deviation (SD) = 10.8). The average Shore value of the infected avocados was 47.3 (SD = 11.8). The average Shore value of the positive control, i.e., the prochloraz-treated avocados was 44.3 (SD = 14.7). The average Shore value of the AITC-treated avocados was 61.1 (SD = 10.5).

[0204] Shore Hardness Measurement: Using Fruit Hardness Tester - HPE III Fff w / Sphere The measurement is performed with a 5.0 mm anvil. The operation process is as follows:

[0205] 1) At room temperature and 1 atmosphere, turn on the measuring device and press the hull or sleeve onto the specimen so that the contact surface of the press plate rests on the fruit. Make sure that the pressure applied to the fruit is sufficient but not so great that the fruit is damaged when the hull is pressed down.

[0206] 2) The results obtained are more reliable if the surface of the skin or peel of the fruit is uniform.

[0207] 3) After placing the hardness tester on the sample, the measurement time starts immediately. The display screen flashes during the measurement.

[0208] 4) Place the measuring device on the fruit until you hear a beep. The measurement is recorded as the Shore hardness of the fruit.

[0209] Example 22. The antifungal activity of avocados exposed to AITC vapor, prochloraz, coating (95 wt% glyceryl monostearate and 5 wt% sodium stearate), coating (95 wt% glyceryl monostearate and 5 wt% sodium stearate) with glyceryl monocaprate, and glyceryl monocaprate was tested against Cg in avocados. To measure the effect of various treatments on ripening, Shore hardness was measured after 5 days of incubation at ambient temperature (about 20°C to about 25°C) and high humidity (about 88% relative humidity to about 93% relative humidity). All samples were wounded by removing the stem and infected by dispensing 1000 Cg spores suspended in 10 μL of water into the wound. Exposure to vapor was accomplished by placing avocados in a closed container, dispensing AITC onto a Whatman filter paper disc, hanging the filter paper disc in front of a battery-powered fan, closing the container and incubating the avocados for 30 minutes. After vapor exposure, the avocados were placed at ambient temperature, high humidity, and evaluated for Cg mold on the 5th day. The test groups consisted of the following avocados: uninfected and untreated; infected and untreated; dipped in a 0.1 g / L prochloraz solution and placed on a rack in front of a fan to dry for 30 minutes; dipped in a 40 g / L solution of the coating agent; dipped in a 3 g / L solution of glyceryl monocaprate; dipped in a 5 g / L solution of glyceryl monocaprate; dipped in a 40 g / L solution of the coating agent and a 3 g / L solution of glyceryl monocaprate; dipped in a 40 g / L solution of the coating agent and a 5 g / L solution of glyceryl monocaprate; exposed to vapor of 125 μL AITC placed on a filter paper disc for 30 minutes; exposed to vapor of 62.5 μL AITC placed on a filter paper disc for 30 minutes; exposed to vapor of 125 μL AITC placed on a filter paper disc for 30 minutes; exposed to vapor of 250 μL AITC placed on a filter paper disc for 30 minutes. and exposed to vapor of 500 μL AITC placed on a filter paper disk for 30 minutes. Fig.29 The Shore hardness results for the test group are shown. The AITC vapor reduced the incidence of Cg mold compared to the untreated sample, the coating agent treated sample, and the prochloraz treated sample. Fig.29 It was shown that the AITC treatment (250 μL) had significantly higher Shores than the uninfected and untreated avocados (average increase of approximately 10 Shores; P value = 0.0016). Fig.29It was shown that the AITC treatment (500 μL) had significantly higher Shores than the uninfected and untreated avocados (average increase of approximately 28.5 Shores; P value < 0.0001).

[0210] Example 23. The ripening characteristics of avocados exposed to AITC vapor, coating agent (95 wt% glyceryl monostearate and 5 wt% sodium stearate), and AITC plus coating agent were analyzed. To measure the effect of various treatments on ripening, Shore hardness was measured after incubation for 11 days at ambient temperature (about 20°C to about 25°C) and high humidity (about 88% relative humidity to about 93% relative humidity). Exposure to vapor was accomplished by placing the avocados in a sealed container, dispensing AITC onto a Whatman filter paper disc, hanging the filter paper disc in front of a battery-powered fan, closing the container and incubating the avocados for 72 hours. The test groups consisted of the following avocados: untreated; immersed in 40 g / L of the coating agent solution; exposed to 500 μL of AITC vapor disposed on a filter paper disc for 30 minutes; immersed in 40 g / L of the coating agent solution, dried, and exposed to 500 μL of AITC vapor disposed on a filter paper disc for 72 hours. Fig. 27 The Shore hardness results for the test groups are shown. The AITC vapor treated avocados were higher quality and had fewer defects than the untreated avocados and the avocados treated with the coating alone. Fig. 27 It was shown that the AITC-treated plus coating test group and the AITC-treated test group had significantly higher Shores than the untreated avocados (mean increase of 30 Shores and 22.5 Shores, respectively; P value < 0.0001). Fig. 27 The coating test group showed slightly higher Shores than the untreated avocados (mean increase of 5.8 Shores; P value < 0.0001).

[0211] Although the present invention includes many specific implementation details, these should not be interpreted as limitations on the scope of the subject matter or on the scope of possible claims, but should be interpreted as descriptions of features that may be specific to some specific implementations. Certain features described in the context of some separate implementations of the present invention may also be implemented in a single implementation in combination. On the contrary, multiple features described in the context of a single implementation may also be implemented in multiple implementations individually or in any suitable sub-combination. In addition, although the features described above may be described as working in certain combinations, even initially claimed as such, in some cases, one or more features in the claimed combination may be deleted from the combination, and the claimed combination may involve a variant of a sub-combination or a sub-combination.

[0212] Specific embodiments of the subject matter have been described. Other embodiments, modifications, and permutations of the described embodiments are within the scope of the following claims and will be apparent to those skilled in the art. Although operations are described in a specific order in the drawings or claims, this should not be construed as requiring that such operations be performed in the specific order or sequential order shown, or that all described operations (some operations may be deemed optional) be performed, in order to obtain the desired results.

[0213] Therefore, the example embodiments described above do not define or constrain the present invention.Other changes, substitutions, and alterations may also be made without departing from the spirit and scope of the present invention.

Claims

1. A method for processing agricultural products, comprising: providing an antimicrobial agent to a housing containing a plurality of produce items; and The plurality of produce items are contacted with the antimicrobial agent, wherein the antimicrobial agent is in gaseous form and is selected to inactivate latent microorganisms in the plurality of produce items.

2. The method of claim 1 , further comprising sealing the enclosure prior to providing the antimicrobial agent to the housing.

3. The method of claim 1, wherein the housing comprises an agricultural product packaging container, an agricultural product storage container, an agricultural product transport container, or an agricultural product processing container.

4. The method of claim 3, wherein the produce packaging container comprises a flip top.

5. The method of claim 3, wherein the produce transport container comprises a shipping container or a box truck.

6. The method of claim 3, wherein the produce processing vessel comprises a chlorosis chamber or a ripening chamber.

7. The method of claim 2, wherein the enclosure is hermetically sealed or restricts the diffusion rate of gas into and out of the enclosure.

8. The method of claim 1, wherein providing the antimicrobial agent to the housing comprises providing a gas flow containing the antimicrobial agent to the housing.

9. The method of claim 8, further comprising pumping the gas flow into the housing.

10. The method of claim 1, wherein the length of time is in the range of 1 minute to 100 hours. The method according to claim 10 , wherein the length of time is in the range of 10 minutes to 1 hour.

12. The method of claim 1, further comprising regulating the temperature or humidity inside the enclosure.

13. The method of claim 1, further comprising volatilizing an antimicrobial agent precursor to produce the antimicrobial agent.

14. The method of claim 13, wherein volatilization of the antimicrobial agent precursor occurs outside of the housing.

15. The method of claim 13, wherein volatilizing the antimicrobial agent precursor occurs within the housing.

16. The method of claim 1, further comprising activating an antimicrobial agent precursor to produce the antimicrobial agent.

17. The method of claim 16, wherein activating the antimicrobial precursor comprises contacting the antimicrobial precursor with a catalyst selected to promote conversion of the antimicrobial precursor to the antimicrobial agent.

18. The method of claim 17, wherein the catalyst is a biocatalyst.

19. The method of claim 18, wherein the biocatalyst is an enzyme.

20. The method of claim 19, wherein the antimicrobial agent precursor comprises alliin, the enzyme comprises alliinase, and the antimicrobial agent comprises allicin.

21. The method of claim 19, wherein the antimicrobial agent comprises sinigrin, the enzyme comprises myrosinase, and the antimicrobial agent comprises allyl isothiocyanate.

22. The method of claim 1, wherein providing the antimicrobial agent to the housing comprises: providing an antimicrobial precursor to a reactor, wherein the reactor comprises a catalyst selected to promote conversion of the antimicrobial precursor to the antimicrobial agent; flowing a carrier gas from a first end of the reactor to a second end of the reactor, thereby forming a gaseous mixture comprising the carrier gas and the antimicrobial agent; and The gaseous mixture is directed from the second end of the fixed bed reactor to the housing.

23. The method of claim 22, wherein providing the antimicrobial precursor to the reactor comprises pumping the antimicrobial precursor toward the reactor.

24. The method of claim 23, wherein flowing the gas from the first end of the reactor to the second end of the reactor comprises forcing the gas to flow from the first end of the reactor to the second end of the reactor.

25. The process of claim 24, wherein the reactor is a fixed bed reactor.

26. The method of claim 24, wherein the catalyst comprises an enzyme.

27. The method of claim 1, wherein contacting the plurality of produce items with the antimicrobial agent comprises circulating the antimicrobial agent within the housing.

28. The method of claim 1, wherein contacting the plurality of produce items with the antimicrobial agent comprises penetrating a skin layer of each of the plurality of produce items with the antimicrobial agent.

29. The method of claim 1, wherein the antimicrobial agent comprises one or more of an antimicrobial agent, an antifungal agent, and an antiviral agent.

30. The method of claim 29, wherein the antimicrobial agent comprises an antifungal agent.

31. The method of claim 30, wherein the antifungal agent comprises allyl isothiocyanate, diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranilate, methyl cinnamate, gamma-decalactone, alpha-terpineol, linalool, or any combination thereof.

32. The method of claim 30, wherein the antifungal agent comprises a derivative of cis-3-hexene.

33. The method of claim 32, wherein the antifungal agent comprises cis-3-hexenol or cis-3-hexenyl acetate.

34. The method of claim 30, wherein the antifungal agent is selected to inactivate green mold or penicillium mold.

35. The method of claim 30, wherein the antifungal agent is selected to inactivate Botrytis cinerea, Penicillium spp., Monilinia spp., Alternaria alternata, Rhizopus stolonifera, Trichothecium roseum, Fusarium spp., Colletotrichum spp., or any combination thereof.

36. The method of claim 1, wherein the plurality of produce items comprises fruits, vegetables, or combinations thereof.

37. The method of claim 36, wherein the plurality of produce items comprises fruit.

38. The method of claim 37, wherein the fruit comprises berries (e.g., avocado, banana, blueberry, cranberry, eggplant, tomato, grape, persimmon), citrus fruits (e.g., orange, lemon, lime, grapefruit, kumquat), gourd fruits (e.g., pumpkin, cucumber, watermelon), stone fruits (e.g., peach, plum, cherry, olive), pome fruits (e.g., apple, pear), aggregate fruits (e.g., blackberry, raspberry), and accessory fruits (e.g., strawberry).

39. The method of claim 1, further comprising placing the plurality of produce items in the enclosure prior to providing the antimicrobial agent to the enclosure.

40. The method of claim 1, further comprising, after contacting the plurality of produce items with the antimicrobial agent for a period of time, removing the plurality of produce items from the enclosure to provide a plurality of treated produce items.

41. The method of claim 1, wherein the antimicrobial agent comprises allyl isothiocyanate.

42. The method of claim 1, wherein the antimicrobial agent comprises allyl isothiocyanate and one or more antimicrobial agents selected from the group consisting of diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranilate, methyl cinnamate, gamma-decalactone, alpha-terpineol, and linalool.

43. The method of claim 41, wherein the antimicrobial agent is provided to the housing in a headspace at a concentration of at least about 1200 ppmv, at least about 1250 ppmv, or from about 1250 ppmv to about 5000 ppmv, or from about 1250 ppmv to about 4900 ppmv.

44. The method of claim 38, wherein the fruit is an avocado, a strawberry, an orange, or a lemon.

45. The method of claim 1, further comprising coating the plurality of produce items with an edible coating composition.

46. ​​The method of claim 45, wherein the edible coating composition comprises: a coating agent comprising one or more saturated glycerides selected from monoglycerides and diglycerides; and one or more fatty acid salts; and Solvent.

47. The method of claim 46, wherein the one or more saturated glycerides are present in the coating agent in an amount of about 75 wt % to about 98 wt %, or about 80 wt % to about 98 wt %, or 85 wt % to about 98 wt %, or about 90 wt % to about 98 wt %, or about 92 wt % to about 97 wt %, or about 95 wt %.

48. The method of claim 46, wherein the one or more fatty acid salts are present in the coating agent in an amount of about 1 wt % to about 20 wt %, or about 1 wt % to about 15 wt %, or about 1 wt % to about 12 wt %, 2 wt % to about 8 wt %, or about 3 wt % to about 7 wt %, or about 5 wt %.

49. The method of claim 46, wherein each of the one or more saturated monoglycerides comprises a carbon chain length of about C10 to about C22.

50. The method of claim 46, wherein one of the one or more saturated glyceryl monoesters is glyceryl monostearate.

51. The method of claim 46, wherein each of the one or more fatty acid salts comprises a carbon chain length of about C10 to about C22.

52. The method of claim 46, wherein one of the fatty acid salts is sodium stearate.

53. The method of claim 46, wherein the coating agent further comprises one or more antimicrobial agents, wherein the one or more antimicrobial agents are selected from diallyl disulfide, carvacrol, eugenol, cinnamaldehyde, limonene, thymol, methyl anthranilate, methyl cinnamate, γ-decanoic acid lactone, α-terpineol, linalool, cis-3-hexenol or cis-3-hexenyl acetate, sodium benzoate, potassium sorbate, chalcone, fludioxonil, 2-hydroxychalcone, 4-hydroxy Chalcone, 4'-hydroxychalcone, 2,2'-dihydroxychalcone, 2,4'-dihydroxychalcone, 2',4-dihydroxychalcone, 2',4'-dihydroxychalcone, 2',4,4'-trihydroxychalcone, 2',4,4'-trihydroxychalcone intermediate, violastyrene, obtusaquinone, apiole, piperine, celastrol, arthonoic acid acid, leoidin, antimycin A, antimycin A1, refractory lichenic acid, ethylorsellinate, methylorsellinate, mycophenolic acid, ethyldichloroorsellinate, angoraxin, isocarpone, eupatoriochromene, xanthoxylum, lichenic acid, aloesin, formononetin, acetovanillin, isoorcellin, deoxysulphurone B 7,4'-dimethyl ether, chrysin dimethyl ether, bergamotol, gambogic acid, 2-hydroxyxanthenone, isoanisole, zanthoxylum lactone, acetylhymetochrome, nobiletin, hymechrome, methoxsalen, 4-methylesculetin, tangerine, kaelin, flavonoids, 3,4',5,6,7-pentamethoxyflavone, deguelin (-), limonene, deoxysappanone B trimethyl ether, deoxysappanone B 7,3'-dimethyl ether, 2',4'-dihydroxy-4-methoxychalcone, daunorubicin hydrochloride, plumbagin, menadione, musciquinone, levomenthol, methyl trimethoxycinnamate, chavicol, cinnamylphenol, benzoate, napthoquinone, benzophenone, acetophenone, benzophenone, phenylacetophenone, chitosan, salicylic acid and sodium salicylate.

54. A system for processing agricultural products, comprising: A reactor configured to facilitate conversion of an antimicrobial agent precursor to an antimicrobial agent, wherein the reactor comprises: a bed configured to contain a particulate media coupled to a catalyst, wherein the catalyst is selected to promote conversion of the antimicrobial agent precursor to the antimicrobial agent; an inlet proximate a first end of the bed and configured to direct a carrier gas from the first end of the bed to a second end of the bed; an additional inlet configured to direct the antimicrobial agent precursor to the bed; and an outlet proximate a second end of the bed and configured to allow a gas mixture comprising the carrier gas and the antimicrobial agent to exit; and A housing is fluidly coupled to the reactor via the outlet, wherein the housing is configured to receive a plurality of produce items and contain a gas mixture in contact with the plurality of produce items.

55. The system of claim 54, wherein the reactor comprises a fixed bed reactor.

56. The system of claim 54, further comprising a pump in fluid communication with the additional inlet and configured to provide the antimicrobial agent precursor to the reactor.

57. The system of claim 56, wherein the pump is a peristaltic pump.

58. The system of claim 54, wherein a carrier gas provided to the inlet volatilizes the antimicrobial agent precursor.

59. A plurality of produce items processed by the method of claim 1.

60. The plurality of produce items of claim 59, wherein the plurality of produce items have a mass loss factor of at least 1.50, at least 1.6, at least 1.7, at least 1.8, at least 1.9, at least 2.0, or at least 2.

5.

61. The plurality of produce items of claim 59, wherein the plurality of produce items are avocados, strawberries, citrus fruits, or lemons.

62. The plurality of produce items of claim 61, wherein the plurality of produce items are 30 treated California #60 avocados, and 5 days after the 30 treated California #60 avocados are treated, the 30 treated California #60 avocados have an average Shore value that is at least 10 Shores, at least 15 Shores, at least 20 Shores, or at least 25 Shores higher than the average Shore value of 30 untreated California #60 avocados, and wherein at day 0, the 30 treated California #60 avocados have an average Shore value that is ±10 Shores of the 30 untreated California #60 avocados.

63. A method of improving the shelf life of a plurality of produce items, the method comprising: providing an antimicrobial agent to a housing containing a plurality of produce items; and The plurality of produce items are contacted with the antimicrobial agent, wherein the antimicrobial agent is in gaseous form and is selected to inactivate latent microorganisms in the plurality of produce items.

64. The method of claim 63, further comprising coating the plurality of produce items with an edible coating composition.

65. A method of delaying the ripening of a plurality of produce items, the method comprising: providing an antimicrobial agent to a housing containing a plurality of produce items; and The plurality of produce items are contacted with the antimicrobial agent, wherein the antimicrobial agent is in gaseous form and is selected to inactivate latent microorganisms in the plurality of produce items.

66. The method of claim 65, further comprising coating the plurality of produce items with an edible coating composition.

67. An edible coating composition comprising: a coating agent comprising: one or more monoglycerides, one or more fatty acid salts, and allyl isothiocyanate; and Solvent.

68. The composition of claim 67, wherein the allyl isothiocyanate is present in the edible coating composition in an amount from about 0.001% v / v to about 5% v / v, or from about 0.01% v / v to about 2.5% v / v, or from about 0.1% v / v to about 1.5% v / v, or from about 0.5% v / v to about 1.5% v / v.

69. The composition of claim 67, wherein the one or more monoglycerides are present in the coating agent in an amount of about 75 wt % to about 98 wt %, about 80 wt % to about 98 wt %, or 85 wt % to about 98 wt %, or about 90 wt % to about 98 wt %, or about 92 wt % to about 97 wt %, or about 95 wt %.

70. The composition of claim 67, wherein the one or more fatty acid salts are present in the coating agent in an amount of about 1 wt % to about 20 wt %, or about 1 wt % to about 15 wt %, or about 1 wt % to about 12 wt %, 2 wt % to about 8 wt %, or about 3 wt % to about 7 wt %, or about 5 wt %.

71. The composition of claim 67, wherein each of the one or more monoglycerides comprises a carbon chain length of about C10 to about C22.

72. The composition of claim 67, wherein one of the one or more saturated monoglycerides is glyceryl monostearate.

73. The composition of claim 67, wherein each of the one or more fatty acid salts comprises a carbon chain length of about C10 to about C22.

74. The composition of claim 67, wherein one of the fatty acid salts is sodium stearate.

75. The composition of claim 67, wherein the solvent comprises water.

76. The composition of claim 67, wherein the coating agent is present in an amount ranging from about 1 g / L to about 150 g / L, or from about 10 g / L to about 100 g / L, or from about 25 g / L to about 75 g / L, or from about 40 g / L to about 60 g / L, or from about 45 g / L to about 55 g / L.

77. A plurality of produce items coated with the edible coating composition of claim 67.

78. The plurality of produce items of claim 77, wherein the plurality of produce items have a mass loss factor of at least 1.50, at least 1.6, at least 1.7, at least 1.8, or at least 2, or at least 2.

5.

79. The plurality of produce items of claim 77, wherein the plurality of produce items are avocados, strawberries, citrus, or lemons.

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