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Title:
LARGE-SCALE METHODS OF UNIFORMLY COATING PACKAGING SURFACES WITH A VOLATILE ANTIMICROBIAL TO PRESERVE FOOD FRESHNESS
Document Type and Number:
WIPO Patent Application WO/2017/180695
Kind Code:
A1
Abstract:
The present application relates to large-scale methods of uniformly coating packaging surfaces with a benzoxaborole compound.

Inventors:
MALEFYT, Timothy (128 Cherry Canyon Drive, Stroudsburg, Pennsylvania, 18360, US)
MACLEAN, Daniel (2574 Allen Circle, Woodland, California, 95776, US)
Application Number:
US2017/027127
Publication Date:
October 19, 2017
Filing Date:
April 12, 2017
Export Citation:
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Assignee:
AGROFRESH INC. (400 Arcola Road, P.O. Box 7000Collegeville, Pennsylvania, 19426, US)
International Classes:
A01N55/08; A23B7/14
Foreign References:
US20150223466A12015-08-13
US20140349853A12014-11-27
US7078546B22006-07-18
US20090148623A12009-06-11
US8110259B22012-02-07
Other References:
EDWARDS: "Organoboron reagents and recent strategies in rhodium catalysed additions", PHD THESIS, 2011, pages 1 - 293, XP055433374
Attorney, Agent or Firm:
FREEMAN, Angela B. (BARNES & THORNBURG LLP, 11 South Meridian StreetIndianapolis, Indiana, 46204, US)
Download PDF:
Claims:
WHAT IS CLAIMED IS:

1. A method of treating a food product with an antimicrobial agent, the method comprising:

administering a benzoxaborole treatment to one or more surfaces of a food packaging material, wherein the benzoxaborole treatment comprises one or more

benzoxaborole compounds,

drying the one or more surfaces of the food packaging material, placing a food product inside of the food packaging material, and vaporizing the benzoxaborole compound from the one or more surfaces of the food packaging material to treat the food product located therein.

2. The method of claim 1, wherein the food product is selected from the group consisting of a strawberry, a raspberry, a blackberry, and a blueberry

3. The method of claim 1, wherein the benzoxaborole compound is

selected from the group consisting of Compound A, Compound B, Compound C, and combinations thereof. 4. The method of claim 3, wherein the benzoxaborole compound is Compound A having the structure:

or an analog or a derivative thereof.

5. The method of claim 3, wherein the benzoxaborole compound is Compound B having the structure: or an analog or a derivative thereof.

6. The method of claim 3, wherein the benzoxaborole compound is Compound C having the structure:

or an analog or a derivative thereof.

7. The method of claim 1, wherein administering the benzoxaborole treatment to food packaging material further comprises embedding the benzoxaborole compound into the food packaging material, impregnating the food packaging material with the benzoxaborole compound, or coating the food packaging material with the benzoxaborole compound.

8. The method of claim 1, wherein the food packaging material is a chamber.

9. The method of claim 8, wherein the chamber is a clamshell.

10. The method of claim 9, wherein the clamshell comprises polyethylene terephthalate.

11. A large-scale method of treating a plurality of chambers with an antimicrobial agent, the method comprising:

placing a plurality of chambers in a position to be treated wherein each of the chambers comprise one or more surfaces,

administering the benzoxaborole treatment to one or more surfaces of the plurality of chambers during preformation, formation, or postformation of the plurality of chambers, wherein the benzoxaborole treatment comprises one or more benzoxaborole compounds,

drying the one or more surfaces of the plurality of chambers, and affixing the benzoxaborole compound to the one or more surfaces of the plurality of chambers.

12. The method of claim 11, wherein the benzoxaborole compound is Compound A having the structure:

or an analog or a derivative thereof.

13. The method of claim 11, wherein the benzoxaborole compound is Compound B having the structure:

or an analog or a derivative thereof.

14. The method of claim 11, wherein the benzoxaborole compound is Compound C having the structure:

or an analog or a derivative thereof.

15. The method of claim 11, wherein administering the benzoxaborole treatment to the one or more surfaces of the plurality of chambers during preformation, formation, or postformation of the plurality of chambers further comprises embedding the benzoxaborole compound into the food packaging material, impregnating the food packaging material with the benzoxaborole compound, or coating the food packaging material with the benzoxaborole compound.

16. The method of claim 11, wherein the one or more surfaces of the

plurality of chambers further comprise a liquid-absorbing material.

17. The method of claim 16, wherein the liquid-absorbing material provides for quick-release or slow-release of the benzoxaborole treatment over a time period.

18. The method of claim 11, wherein the plurality of chambers are a plurality of clamshells.

19. The method of claim 18, wherein the plurality of clamshells comprise polyethylene terephthalate.

20. The method of claim 11, wherein drying the one or more surfaces of the plurality of chambers occurs instantaneously.

Description:
LARGE-SCALE METHODS OF UNIFORMLY COATING PACKAGING SURFACES WITH A VOLATILE ANTIMICROBIAL TO PRESERVE FOOD FRESHNESS

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the benefit under 35 USC § 119(e) of U.S. Provisional Patent Application Serial No. 62/323,247, filed on April 15, 2016, the entire disclosure of which is incorporated herein by reference.

FIELD OF THE PRESENT APPLICATION

The present application relates to large-scale methods of uniformly coating packaging surfaces with a benzoxaborole compound.

BACKGROUND

Benzoxaborole is a drug known to be effective in treating eukaryotic fungal

and parasitic infections. For example, benzoxaborole is used to treat fungal conditions affecting the toenails and fingernails of humans, such as Onychomycosis. Benzoxaborole is also known to be an effective treatment of Human African Trypanosomiasis, commonly called African Sleeping Sickness, which is caused by T. brucei parasites that infect thousands of people annually in sub-Saharan Africa.

Benzoxaborole has also been shown to have antimicrobial effects in plants.

For example, benzoxaborole compounds have been proven to be effective as a volatile plant fungicide. However, coating packaging surfaces with a volatile biological control agent, such as a benzoxaborole compound, to provide antimicrobial protection to food, such as crops and meats, and to preserve food freshness has not been reported.

The present disclosure describes large-scale methods of applying volatile antimicrobial compounds on food containers and packaging materials to inhibit microorganisms responsible for decay. More specifically, the present disclosure provides methods of coating

benzoxaboroles on the surface of food containers and packaging materials in mass in order to provide antimicrobial protection to food. In addition, the present disclosure describes methods to treat food products against microorganisms that are detrimental to the preservation of the freshness of food, such as plant, crops, or meats.

SUMMARY OF THE INVENTION

The present disclosure provides a method of treating food products with an

antimicrobial agent. The method comprises administering a benzoxaborole compound to one or more surfaces of a chamber. The method also comprises drying the one or more surfaces of the chamber. The method further comprises placing a food product inside of the chamber. Finally, the method comprises vaporizing the benzoxaborole compound from the one or more surfaces of the chamber to treat the food product.

In the method described herein, the food product may be a plant crop or a meat, wherein the crop may be a fruit selected from the group consisting of a strawberry, a raspberry, a blackberry, or a blueberry. In addition, the benzoxaborole compound of the present method may be selected from the group consisting of Compound A, Compound B, and/or Compound C. The chamber of the method may be a clamshell, and the clamshell may comprise polyethylene terephthalate. Finally, the benzoxaborole compound of the present method may be

administered to the chamber by drenching, spraying, painting, vaporizing, and/or sublimation.

The present disclosure is also directed to a large-scale method of treating a plurality of chambers with an antimicrobial agent. The method comprises placing a plurality of chambers in a position to be treated, wherein each of the plurality of chambers comprises one or more surfaces. The method further comprises administering a benzoxaborole compound to the one or more surfaces of the plurality of chambers. In addition, the method comprises drying the one or more surfaces of the plurality of chambers, wherein the drying may be instantaneous. Further, the large-scale method comprises affixing the benzoxaborole compound to the one or more surfaces of the plurality of chambers.

The benzoxaborole compound of the present large-scale method may be selected from the group consisting of Compound A, Compound B, and/or Compound C, and may be administered to the plurality of chambers by drenching, spraying, painting, vaporizing, or sublimation. The benzoxaborole compound may also be administered

to the plurality of chambers during preformation, formation, or postformation of the plurality of chambers. The surfaces of the plurality of chambers of the large-scale method may further comprise a liquid-absorbing material. The plurality of chambers of the large-scale method may comprise a plurality of clamshells, and the plurality of clamshells may comprise polyethylene terephthalate. DETAILED DESCRIPTION

The following numbered embodiments are contemplated and are non-limiting:

1. A method of treating a food product with an antimicrobial agent, the

method comprising:

administering a benzoxaborole treatment to one or more surfaces of a food packaging material, wherein the benzoxaborole treatment comprises one or more benzoxaborole compounds,

drying the one or more surfaces of the food packaging material,

placing a food product inside of the food packaging material, and

vaporizing the benzoxaborole compound from the one or more surfaces of the food packaging material to treat the food product located therein.

2. The method of clause 1, wherein the food product is a plant, a crop, or a meat. 3. The method of clause 1 or clause 2, wherein the food product is a fruit.

4. The method of any of clauses 1 to 3, wherein the food product is selected from the group consisting of a strawberry, a raspberry, a blackberry, and a blueberry. 5. The method of any one of clauses 1 to 4, wherein the benzoxaborole compound is selected from the group consisting of Compound A, Compound B, Compound C, and combinations thereof.

6. The method of any one of clauses 1 to 5, wherein the benzoxaborole compound is Compound A having the structure:

or an analog or a derivative thereof.

7. The method of any one of clauses 1 to 5, wherein the benzoxaborole compound is Compound B having the structure: or an analog or a derivative thereof.

8. The method of any one of clauses 1 to 5, wherein the benzoxaborole compound is Compound C having the structure:

or an analog or a derivative thereof.

9. The method of any one of clauses 1 to 8, wherein the benzoxaborole compound is in the form of a liquid, a gas, or a solid.

10. The method of any one of clauses 1 to 9, wherein the food packaging material is comprised by a chamber.

11. The method of clause 10, wherein the chamber is selected from the group consisting of a container, a liner material, one chamber, and a plurality of chambers.

12. The method of clause 10 or clause 11, wherein the chamber is a clamshell.

13. The method of clause 12, wherein the clamshell comprises polyethylene terephthalate (PET).

14. The method of any one of clauses 1 to 13, wherein the benzoxaborole treatment is embedded into, impregnated within, or coated onto the food packaging material. 15. The method of any one of clauses 1 to 14, wherein the benzoxaborole treatment is embedded into the food packaging material.

16. The method of any one of clauses 1 to 14, wherein the benzoxaborole treatment is coated onto the food packaging material.

17. The method of any one of clauses 1 to 14, wherein the benzoxaborole treatment is impregnated into the food packaging material. 18. The method of any one of clauses 1 to 17, wherein the benzoxaborole treatment is in the form of a spray, a liquid, a mist, a gel, a thermal fog, a non-thermal fog, a dip, a drench, a vapor, a gas, or sublimation.

19. The method of any one of clauses 1 to 18, wherein the benzoxaborole treatment further comprises a treatment carrier.

20. The method of clause 19, wherein the treatment carrier comprises a liquid, a gas, a solution, a solvent, and a chemical. 21. The method of clause 19 or clause 20, wherein the treatment carrier is a liquid.

22. The method of any one of clauses 19 to 21, wherein the treatment carrier is selected from the group consisting of water, saline, a buffer, a solution, a solvent, a solvent- based solution, and a combination thereof.

23. The method of clause any one of clauses 19 to 22, wherein the treatment carrier is supercritical C0 2 .

24. The method of clause 19 or clause 20, wherein the treatment carrier is a gas.

25. The method of any one of clauses 19, 20, or 24, wherein the treatment carrier is selected from the group consisting of nitrogen (N 2 ), carbon dioxide (C0 2 ), and sulfur dioxide (S0 2 ). 26. The method of any one of clauses 19, 20, 24, or 25, wherein the treatment carrier is nitrogen (N 2 ).

27. The method of any one of clauses 19, 20, 24, or 25, wherein the treatment carrier is carbon dioxide (C0 2 ).

28. The method of any one of clauses 19, 20, 24, or 25, wherein the treatment carrier is sulfur dioxide (S0 2 ). 29. The method of any one of clauses 1 to 28, wherein the benzoxaborole treatment is effective against plant pathogens.

30. The method of clause 29, wherein the plant pathogens are fungal pathogens. 31. The method of clause 29 or clause 30, wherein the plant pathogens are selected from the group consisting of Acremonium spp., Albugo spp., Alternaria spp., Ascochyta spp., Aspergillus spp., Botryodiplodia spp., Botryospheria spp., Botrytis spp., Byssochlamys spp., Candida spp., Cephalosporium spp., Ceratocystis spp., Cercospora spp., Chalara spp., Cladosporium spp., Colletotrichum spp., Cryptosporiopsis spp., Cylindrocarpon spp.,

Debaryomyces spp., Diaporthe spp., Didymella spp., Diplodia spp., Dothiorella spp., Elsinoe spp., Fusarium spp., Geotrichum spp., Gloeosporium spp., Glomerella spp., Helminthosporium spp., Khuskia spp., Lasiodiplodia spp., Macrophoma spp., Macrophomina spp., Microdochium spp., Monilinia spp., Monilochaethes spp., Mucor spp., Mycocentrospora spp., Mycosphaerella spp., Nectria spp., Neofabraea spp., Nigrospora spp., Penicillium spp., Peronophythora spp., Peronospora spp., Pestalotiopsis spp., Pezicula spp., Phacidiopycnis spp., Phoma spp.,

Phomopsis spp., Phyllosticta spp., Phytophthora spp., Polyscytalum spp., Pseudocercospora spp., Pyricularia spp., Pythium spp., Rhizoctonia spp., Rhizopus spp., Sclerotium spp.,

Sclerotinia spp., Septoria spp., Sphaceloma spp., Sphaeropsis spp., Stemphyllium spp., Stilbella spp., Thielaviopsis spp., Thyronectria spp., Trachysphaera spp., Uromyces spp., Ustilago spp., Venturia spp., and Verticillium spp., and bacterial pathogens, such as Bacillus spp.,

Campylobacter spp., Clavibacter spp., Clostridium spp., Erwinia spp., Escherichia spp., Lactobacillus spp., Leuconostoc spp., Listeria spp., Pantoea spp., Pectobacterium spp., Pseudomonas spp., Ralstonia spp., Salmonella spp., Shigella spp., Staphylococcus spp., Vibrio spp., Xanthomonas spp., and Yersinia spp. 32. The method of any one of clauses 29 to 31, wherein the plant pathogens are selected from the group consisting of Botrytis cinerea, Mucor piriformis, Fusarium

sambucinum, Aspergillus brasiliensis, and Peniciliium expansum. 33. The method of any one of clauses 10 to 32, wherein the chamber is open, closed, or sealed.

34. The method of any one of clauses 10 to 33, wherein the chamber is sealed. 35. The method of any one of clauses 10 to 34, wherein the chamber is air-tight.

36. The method of any one of clauses 10 to 35, wherein the chamber is

semipermeable or impermeable. 37. The method of any one of clauses 10 to 36, wherein the chamber is made of a material selected from the group consisting of cardboard, paper, paperboard, corrugated paper, plastic, glass, polyester, polystyrene, cellulosic material, metal, and cement.

38. The method of clause 37, wherein the metal is selected from the group consisting of aluminum, foils, laminates, tinplate, and steel.

39. The method of clause 38, wherein the steel is tin-free steel.

40. The method of clause 37, wherein the plastic is selected from the group consisting of thermosets and thermoplastics.

41. The method of clause 37, wherein the polyester is selected from the group consisting of polycarbonate, polyethylene naphthalate, and polyethylene terephthalate (PET). 42. The method of any one of clauses 10 to 41, wherein the chamber comprises a port, an outlet, or both.

43. The method of any one of clauses 10 to 42, wherein the chamber has a volume from between about 0.1 L to about 50 L. 44. The method of any one of clauses 10 to 43, wherein the chamber comprises a plurality of individual clamshells.

45. The method of any one of clauses 10 to 44, wherein the chamber comprises between about 2 clamshells to about 384,000,000 clamshells.

46. The method of any one of clauses 10 to 45, wherein the chamber further comprises a liquid- absorbing material. 47. The method of clause 46, wherein the liquid-absorbing material is comprised on the interior of the chamber or on the exterior of the chamber.

48. The method of clause 46 or clause 47, wherein the liquid- absorbing material is comprised on the interior of the chamber.

49. The method of clause 47 or clause 48, wherein the liquid- absorbing material is comprised on the internal top, the internal bottom, or the internal side panels of the chamber.

50. The method of clause 46 or clause 47, wherein the liquid- absorbing material is comprised on the exterior of the chamber.

51. The method of clause 47 or clause 50, wherein the liquid- absorbing material is comprised on the external top, the external bottom, or the external side panels of the chamber. 52. The method of any one of clauses 46 to 51, wherein the liquid- absorbing material is attached or affixed to the chamber.

53. The method of any one of clauses 46 to 52, wherein the liquid- absorbing material is comprised in a chamber component selected from the group consisting of a liner, a wrapping, a label, a tag, a sticker, and a pad.

54. The method of any one of clauses 45 to 53, wherein the liquid- absorbing material is selected from the group consisting of cotton, paper, and foam. 55. The method of any one of clauses 46 to 54, wherein the liquid- absorbing material is a reservoir capable of releasing the benzoxaborole treatment to the food product comprised in the chamber.

56. The method of any one of clauses 46 to 55, wherein the liquid- absorbing material provides for quick-release or slow-release of the benzoxaborole treatment to the food product comprised in the chamber over a time period.

57. The method of any one of clauses 46 to 56, wherein the liquid- absorbing material provides for quick-release of the benzoxaborole treatment to the food product comprised in the chamber over a time period.

58. The method of any one of clauses 46 to 56, wherein the liquid- absorbing material provides for slow-release of the benzoxaborole treatment to the food product comprised in the chamber over a time period.

59. The method of clause 56 or clause 57, wherein the time period for quick-release of the benzoxaborole treatment by the liquid-absorbing material is about 12 hours or less. 60. The method of any one of clauses 56, 57, or 59, wherein the time period for quick-release of the benzoxaborole treatment by the liquid- absorbing material is between about 5 seconds to about 12 hours or less.

61. The method of clause 56 or clause 58, wherein the time period for slow-release of the benzoxaborole treatment by the liquid-absorbing material is over 12 hours.

62. The method of any one of clauses 56, 58, or 61, wherein the time period for slow-release of the benzoxaborole treatment by the liquid-absorbing material is between over 12 hours to about 31 days.

63. The method of any one of clauses 10 to 62, wherein the chamber further comprises one or more apertures.

64. The method of clause 63, wherein the one or more apertures has a size between about 2 mm to about 2 cm. 65. The method of clause 63 or clause 64, wherein the one or more apertures has a location on the chamber selected from the group consisting of the base, the lid, the sides, or a combination thereof. 66. The method of any one of clauses 63 to 65, wherein the one or more apertures enables introduction of the benzoxaborole treatment, the treatment carrier, or a combination thereof to the chamber.

67. The method of any one of clauses 63 to 65, wherein the one or more apertures enables release of the benzoxaborole treatment, the treatment carrier, or a combination thereof from the chamber.

68. The method of any one of clauses 1 to 67, wherein the food products are manually or robotically placed in the chamber.

69. The method of any one of clauses 1 to 68, wherein the food products are treated post-harvest.

70. The method of any one of clauses 1 to 69, wherein the distance between the food products and the food packaging material comprising the benzoxaborole treatment is no greater than 6 feet.

71. The method of any one of clauses 1 to 70, wherein the distance between the food products and the food packaging material comprising the benzoxaborole treatment is between about 0.1 inches and about 6 feet.

72. The method any one of clauses 1 to 71, wherein the one or more food products are treated for an initial time period ranging from about 12 hours to about 5 days. 73. The method of any one of clauses 1 to 72, wherein the benzoxaborole treatment concentration ranges from about 0.1 mg/chamber to about 10 mg/chamber.

74. The method of any one of clauses 1 to 73, wherein drying the one or more surfaces of the food packaging material occurs at room temperature, wherein room temperature is between about 21°C and about 23 °C. 75. The method of any one of clauses 1 to 74, wherein drying the one or more surfaces of the food packaging material occurs instantaneously or within seconds.

76. The method of any one of clauses 1 to 75, wherein drying the one or more surfaces of the food packaging material occurs instantaneously.

77. The method of any one of clauses 1 to 75, wherein drying the one or more surfaces of the food packaging material occurs within seconds. 78. The method of any one of clauses 1 to 75 or 77, wherein drying the one or more surfaces of the food packaging material occurs between about 0.1 seconds and about 60 seconds.

79. The method of any one of clauses 1 to 78, wherein the method results in greater uniformity and consistency in application of benzoxaborole treatment to the food packaging material.

80. The method of any one of clauses 1 to 79, wherein the method provides one month of extended antimicrobial protection to the treated food products.

81. A large-scale method of treating a plurality of chambers with an

antimicrobial agent, the method comprising:

placing a plurality of chambers in a position to be treated wherein each of the chambers comprise one or more surfaces,

administering the benzoxaborole treatment to one or more surfaces of the plurality of chambers during preformation, formation, or postformation of the plurality of chambers, wherein the benzoxaborole treatment comprises one or more benzoxaborole compounds,

drying the one or more surfaces of the plurality of chambers, and affixing the benzoxaborole compound to the one or more surfaces of the plurality of chambers.

82. The method of clause 81, wherein the plurality of chambers further

comprise a food product that is a plant, a crop, or a meat. 83. The method of clause 82, wherein the food product is a fruit.

84. The method of any of clauses 82 or clause 83, wherein the food product is selected from the group consisting of a strawberry, a raspberry, a blackberry, and a blueberry.

85. The method of any one of clauses 81 to 84, wherein the benzoxaborole compound is selected from the group consisting of Compound A, Compound B, Compound C, and combinations thereof.

86. The method of any one of clauses 81 to 85, wherein the benzoxaborole compound is Compound A having the structure:

or an analog or a derivative thereof.

87. The method of any one of clauses 81 to 85, wherein the benzoxaborole compound is Compound B having the structure:

or an analog or a derivative thereof. 88. The method of any one of clauses 81 to 85, wherein the benzoxaborole compound is Compound C having the structure: or an analog or a derivative thereof.

89. The method of any one of clauses 81 to 88, wherein the benzoxaborole compound is in the form of a liquid, a gas, or a solid.

90. The method of any one of clauses 81 to 89, wherein the plurality of chambers comprise one or more containers. 91. The method of any one of clauses 81 to 90, wherein the plurality of chambers comprise one or more liner materials.

92. The method of any one of clauses 81 to 91, wherein the plurality of chambers comprise one or more clamshells.

93. The method of any one of clauses 81 to 92, wherein the clamshells comprise polyethylene terephthalate (PET).

94. The method of any one of clauses 81 to 93, wherein the benzoxaborole treatment is embedded into, impregnated within, or coated onto the plurality of chambers.

95. The method of any one of clauses 81 to 94, wherein the benzoxaborole treatment is embedded into the plurality of chambers. 96. The method of any one of clauses 81 to 94, wherein the benzoxaborole treatment is coated onto the plurality of chambers.

97. The method of any one of clauses 81 to 94, wherein the benzoxaborole treatment is impregnated into the plurality of chambers. 98. The method of any one of clauses 81 to 97, wherein the benzoxaborole treatment is in the form of a spray, a liquid, a mist, a gel, a thermal fog, a non-thermal fog, a dip, a drench, a vapor, a gas, or sublimation. 99. The method of any one of clauses 81 to 98, wherein the benzoxaborole treatment further comprises a treatment carrier.

100. The method of clause 99, wherein the treatment carrier comprises a liquid, a gas, a solution, a solvent, and a chemical.

101. The method of clause 99 or clause 100, wherein the treatment carrier is a liquid.

102. The method of any one of clauses 99 to 101, wherein the treatment carrier is selected from the group consisting of water, saline, a buffer, a solution, a solvent, a solvent- based solution, and a combination thereof.

103. The method of clause any one of clauses 99 to 102, wherein the treatment carrier is supercritical C0 2 . 104. The method of clause 99 or clause 100, wherein the treatment carrier is a gas.

105. The method of any one of clauses 99, 100, or 104, wherein the treatment carrier is selected from the group consisting of nitrogen (N 2 ), carbon dioxide (C0 2 ), and sulfur dioxide (S0 2 ).

106. The method of any one of clauses 99, 100, 104, or 105, wherein the treatment carrier is nitrogen (N 2 ).

107. The method of any one of clauses 99, 100, 104, or 105, wherein the treatment carrier is carbon dioxide (C0 2 ).

108. The method of any one of clauses 99, 100, 104, or 105, wherein the treatment carrier is sulfur dioxide (S0 2 ). 109. The method of any one of clauses 81 to 108, wherein the benzoxaborole treatment is effective against plant pathogens.

110. The method of clause 109, wherein the plant pathogens are fungal pathogens.

111. The method of clause 109 or clause 110, wherein the plant pathogens are selected from the group consisting of Acremonium spp., Albugo spp., Alternaria spp., Ascochyta spp., Aspergillus spp., Botryodiplodia spp., Botryospheria spp., Botrytis spp., Byssochlamys spp., Candida spp., Cephalosporium spp., Ceratocystis spp., Cercospora spp., Chalara spp., Cladosporium spp., Colletotrichum spp., Cryptosporiopsis spp., Cylindrocarpon spp., Debaryomyces spp., Diaporthe spp., Didymella spp., Diplodia spp., Dothiorella spp., Elsinoe spp., Fusarium spp., Geotrichum spp., Gloeosporium spp., Glomerella spp.,

Helminthosporium spp., Khuskia spp., Lasiodiplodia spp., Macrophoma spp., Macrophomina spp., Microdochium spp., Monilinia spp., Monilochaethes spp., Mucor spp., Mycocentrospora spp., Mycosphaerella spp., Nectria spp., Neofabraea spp., Nigrospora spp., Penicillium spp., Peronophythora spp., Peronospora spp., Pestalotiopsis spp., Pezicula spp., Phacidiopycnis spp., Phoma spp., Phomopsis spp., Phyllosticta spp., Phytophthora spp., Polyscytalum spp., Pseudocercospora spp., Pyricularia spp., Pythium spp., Rhizoctonia spp., Rhizopus spp., Sclerotium spp., Sclerotinia spp., Septoria spp., Sphaceloma spp., Sphaeropsis spp.,

Stemphyllium spp., Stilbella spp., Thielaviopsis spp., Thyronectria spp., Trachysphaera spp., Uromyces spp., Ustilago spp., Venturia spp., and Verticillium spp., and bacterial pathogens, such as Bacillus spp., Campylobacter spp., Clavibacter spp., Clostridium spp., Erwinia spp., Escherichia spp., Lactobacillus spp., Leuconostoc spp., Listeria spp., Pantoea spp.,

Pectobacterium spp., Pseudomonas spp., Ralstonia spp., Salmonella spp., Shigella spp., Staphylococcus spp., Vibrio spp., Xanthomonas spp., and Yersinia spp.

112. The method of any one of clauses 109 to 111, wherein the plant pathogens are selected from the group consisting of Botrytis cinerea, Mucor piriformis, Fusarium

sambucinum, Aspergillus brasiliensis, and Peniciliium expansum.

113. The method of any one of clauses 90 to 112, wherein the chamber is open, closed, or sealed.

114. The method of clause 113, wherein the chamber is sealed. 115. The method of clause 113 or clause 114, wherein the chamber is sealed air-tig

116. The method of any one of clauses 90 to 115, wherein the chamber is semipermeable or impermeable.

117. The method of any one of clauses 90 to 116, wherein the chamber is made of a material selected from the group consisting of cardboard, paper, paperboard, corrugated paper, plastic, glass, polyester, polystyrene, cellulosic material, metal, and cement. 118. The method of clause 117, wherein the metal is selected from the group consisting of aluminum, foils, laminates, tinplate, and steel.

119. The method of clause 118, wherein the steel is tin-free steel. 120. The method of clause 117, wherein the plastic is selected from the group consisting of thermosets and thermoplastics.

121. The method of clause 117, wherein the polyester is selected from the group consisting of polycarbonate, polyethylene naphthalate, and polyethylene terephthalate (PET).

122. The method of any one of clauses 90 to 121, wherein the chamber comprises a port, an outlet, or both.

123. The method of any one of clauses 81 to 122, wherein the chamber has a volume from between about 0.1 L to about 50 L.

124. The method of any one of clauses 81 to 123, wherein the chamber comprises a plurality of individual clamshells. 125. The method of any one of clauses 81 to 124, wherein the chamber comprises between about 2 clamshells to about 384,000,000 clamshells.

126. The method of any one of clauses 81 to 125, wherein the chamber further comprises a liquid- absorbing material. 127. The method of clause 126, wherein the liquid-absorbing material is comprised on the interior of the chamber or on the exterior of the chamber.

128. The method of clause 126 or clause 127, wherein the liquid- absorbing material is comprised on the interior of the chamber.

129. The method of clause 127 or clause 128, wherein the liquid- absorbing material is comprised on the internal top, the internal bottom, or the internal side panels of the chamber, or combinations thereof.

130. The method of clause 126 or clause 127, wherein the liquid- absorbing material is comprised on the exterior of the chamber.

131. The method of clause 127 or clause 130, wherein the liquid- absorbing material is comprised on the external top, the external bottom, or the external side panels of the chamber, or combinations thereof.

132. The method of any one of clauses 126 to 131, wherein the liquid- absorbing material is attached or affixed to the chamber.

133. The method of any one of clauses 126 to 132, wherein the liquid- absorbing material is comprised in a chamber component selected from the group consisting of a liner, a wrapping, a label, a tag, a sticker, and a pad. 134. The method of any one of clauses 126 to 133, wherein the liquid- absorbing material is selected from the group consisting of cotton, paper, and foam.

135. The method of any one of clauses 126 to 134, wherein the liquid- absorbing material is a reservoir capable of releasing the benzoxaborole treatment to the food product comprised in the chamber.

136. The method of any one of clauses 126 to 135, wherein the liquid- absorbing material provides for quick-release or slow-release of the benzoxaborole treatment to the food product comprised in the chamber over a time period. 137. The method of any one of clauses 126 to 136, wherein the liquid- absorbing material provides for quick-release of the benzoxaborole treatment to the food product comprised in the chamber over a time period. 138. The method of any one of clauses 126 to 136, wherein the liquid- absorbing material provides for slow-release of the benzoxaborole treatment to the food product comprised in the chamber over a time period.

139. The method of clause 136 or clause 137, wherein the time period for quick- release of the benzoxaborole treatment by the liquid-absorbing material is about 12 hours or less.

140. The method of any one of clauses 136, 137, or 139, wherein the time period for quick-release of the benzoxaborole treatment by the liquid- absorbing material is between about 5 seconds to about 12 hours or less.

141. The method of clause 136 or clause 138, wherein the time period for slow- release of the benzoxaborole treatment by the liquid-absorbing material is over 12 hours. 142. The method of any one of clauses 136, 138, or 141, wherein the time period for slow-release of the benzoxaborole treatment by the liquid-absorbing material is between over 12 hours to about 31 days.

143. The method of any one of clauses 81 to 142, wherein the plurality of chambers further comprises one or more apertures.

144. The method of clause 143, wherein the one or more apertures has a size between about 2 mm to about 2 cm. 145. The method of clause 143 or clause 144, wherein the one or more apertures is in a location on the chamber selected from the group consisting of the base, the lid, the sides, or a combination thereof. 146. The method of any one of clauses 143 to 145, wherein the one or more apertures enables introduction of the benzoxaborole treatment, the treatment carrier, or a combination thereof to the chamber. 147. The method of any one of clauses 143 to 145, wherein the one or more apertures enables release of the benzoxaborole treatment, the treatment carrier, or a combination thereof from the chamber.

148. The method of any one of clauses 81 to 147, wherein the food products are manually or robotically placed in the chamber.

149. The method of any one of clauses 81 to 148, wherein the food products are treated post-harvest. 150. The method of any one of clauses 81 to 149, wherein the distance between the food products and the plurality of chambers comprising the benzoxaborole treatment is no greater than 6 feet.

151. The method of any one of clauses 81 to 150, wherein the distance between the food products and the plurality of chambers comprising the benzoxaborole treatment is between about 0.1 inches and about 6 feet.

152. The method any one of clauses 81 to 151, wherein the one or more food products are treated for an initial time period ranging from about 12 hours to about 5 days.

153. The method of any one of clauses 81 to 152, wherein the benzoxaborole treatment concentration ranges from about 0.1 mg/chamber to about 10 mg/chamber.

154. The method of any one of clauses 81 to 153, wherein drying the one or more surfaces of the plurality of chambers occurs at room temperature, wherein room temperature is between about 21°C and about 23 °C.

155. The method of any one of clauses 81 to 154, wherein drying the one or more surfaces of the plurality of chambers occurs instantaneously or within seconds. 156. The method of any one of clauses 81 to 155, wherein drying the one or more surfaces of the plurality of chambers occurs instantaneously.

157. The method of any one of clauses 81 to 155, wherein drying the one or more surfaces of the plurality of chambers occurs within seconds.

158. The method of any one of clauses 81 to 155 or 157, wherein drying the one or more surfaces of the plurality of chambers occurs between about 0.1 seconds and about 60 seconds.

159. The method of any one of clauses 81 to 158, wherein the method results in greater uniformity and consistency in application of benzoxaborole treatment to the plurality of chambers.

160. The method of any one of clauses 81 to 159, wherein the method provides one month of extended antimicrobial protection to the treated food products.

The terms "chamber," "container," "material," or the phrase "packaging material" are interchangeable and refer to any material that is used to box, wrap, store, or package a food or food product, such as a plant, crop, or meat. A plurality of chambers comprises from about 1000 to tens of thousands to one or more millions of chambers.

The term "carrier" refers to a material, composition, or control, such as a liquid or solid filler, diluent, excipient, solvent, gas, or encapsulating material, involved in carrying or transporting an active ingredient, compound, analog, or derivative from one location to another location. A carrier must be acceptable in the sense of being compatible with the other ingredients of the formulation and not injurious to food, such as plant, crops, or meat products.

Exemplary embodiments of the compounds of the present disclosure comprise

Compounds A, B, and/or C, which may encompass diastereomers and enantiomers of the illustrative compounds. Enantiomers are defined as one of a pair of molecular entities which are mirror images of each other and non-superimposable. Diastereomers or diastereoisomers are defined as stereoisomers other than enantiomers. Diastereomers or diastereoisomers are stereoisomers not related as mirror images. Diastereoisomers are characterized by differences in physical properties.

The terms "food" or "food product" refer to a plant or plant parts.

The term "plant(s)" and "plant parts" include, but not limited to, plant tissues, such as leaves, calli, stems, roots, flowers, fruits, vegetables, pollen, and seeds. A class of plants that may be used in the present invention is generally as broad as the class of higher and lower plants including, but not limited to, dicotyledonous plants, monocotyledonous plants, and plant crops, including, but not limited to, vegetable crops, fruit crops, ornamental crops, and meats.

"Vegetable crops" include, but are not limited to, asparagus, beet (e.g., sugar beet and fodder beet), beans, broccoli, cabbage, carrot, cassava, cauliflower, celery, cucumber, eggplant, garlic, gherkin, leafy greens (lettuce, kale, spinach, and other leafy greens), leek, lentils, mushroom, onion, peas, pepper (e.g., sweet peppers, bell peppers, and hot peppers), potato, pumpkin, sweet potato, snap bean, squash, tomato, and turnip.

"Fruit crops" include, but are not limited to, apple, avocado, banana, soft fruits, such as, strawberry, blueberry, raspberry, blackberry, cranberry, currents and other types of soft fruit berries, carambola, cherry, citrus (e.g., oranges, lemon, lime, mandarin, grapefruit, and other citrus), coconut, fig, grapes, guava, kiwifruit, mango, nectarine, melons (including cantaloupe, muskmelon, watermelon, and other melons), olive, papaya, passionfruit, peach, pear, persimmon, pineapple, plum, and pomegranate. More specifically, horticultural crops of the present disclosure include, but are not limited to, soft fruits (e.g., grape, apple, pear, and persimmon) and berries (e.g., strawberries, blackberries, blueberries, and raspberries).

"Ornamental crops" include, but are not limited to, baby's breath, carnation, dahlia, daffodil, geranium, gerbera, lily, orchid, peony, Queen Anne's lace, rose, snapdragon, or other cut-flowers or ornamental flowers, potted flowers, flower bulbs, shrub, and deciduous or coniferous tree.

"Meat" or "Meats" include, but are not limited to beef, bison, chicken, deer, goat, turkey, pork, sheep, fish, shellfish, mollusks, or dry-cured meat products.

The term "subliming" refers the ability of a chemical, compound, or composition or other solid substance to evaporate or to disperse into vapor or gas when heated. Often the substance can transition back to a solid upon cooling.

The term "vaporizing" refers to transitioning or converting into vapor.

The term "volatile" or "volatilizes" refers to the ability of a chemical, compound, or composition or other substance to evaporate or to disperse into vapor or gas.

COMPOUNDS AND COMPONENTS OF THE PRESENT METHODS

The methods of the present disclosure are directed to treating food packaging materials or containers with one or more volatile antimicrobial compounds. More specifically, the methods described herein provide for coating benzoxaborole compounds on surfaces of food packaging materials or containers in order to delay or inhibit microorganism growth and food decay. Accordingly, the methods of the present disclosure comprise, consist of, or consist essentially of benzoxaborole compounds. Exemplary embodiments of the compounds of the present disclosure comprise

Compounds A, B, and C, which may encompass diastereomers and enantiomers of the illustrative compounds. Enantiomers are defined as one of a pair of molecular entities which are mirror images of each other and non-superimposable. Diastereomers or diastereoisomers are defined as stereoisomers other than enantiomers. Diastereomers or diastereoisomers are stereoisomers not related as mirror images. Diastereoisomers are characterized by differences in physical properties.

One exemplary embodiment of a benzoxaborole compound of the present method is Compound A:

or an analog or derivative thereof. An additional illustrative embodiment of a benzoxaborole compound of the present method is Compound B:

or an analog or derivative thereof.

Another exemplary embodiment of a benzoxaborole compound of the present method is Compound C, which is a salt version of Compounds A and/or B:

or an analog or derivative thereof.

Compounds A, B, and/or C may be used individually or as a mixture or combination. The benzoxaborole compounds may also be used in combination with a carrier to form a benzoxaborole treatment. The benzoxaborole treatment provides antimicrobial protection to food, such as plants, crops, or meats, when administered, applied, or exposed to the plants, crops, or meats. While the mechanism of action of the benzoxaborole compound is not fully understood, it is thought to proceed via blocking or inhibition of protein synthesis in microorganisms and/or blocking cytoplasmic leucyl-tRNA synthetase (LeuRS) activity, thereby preventing growth of microorganisms on food.

Benzoxaborole Compounds A, B, and/or C may be used in any form, including, but not limited to, a liquid, a solid, or a gaseous composition. In particular, the present method provides application of a benzoxaborole compound to food product packaging materials as a spray, a mist, a gel, a thermal and non-thermal fog, a dip, a drench, a vapor, a gas, or via sublimation.

Carriers of the present disclosure may be combined with one or more active

benzoxaborole compounds to form a benzoxaborole treatment. Treatment carriers of the present disclosure may comprise gases, solutions, solvents, or chemicals. For example, a liquid carrier of the present disclosure may comprise water, buffer, saline solution, a solvent, or solvent-based solution, etc. Illustrative liquid solvent carriers of the present disclosure include, but are not limited to, liquid carbon dioxide (C0 2 ), such as supercritical C0 2 . Gaseous carriers of the the benzoxaborole compounds may comprise nitrogen (N 2 ), carbon dioxide (C0 2 ), or sulfur dioxide (S0 2 ).

Benzoxaborole compound treatments comprising Compounds A, B, and/or C, with or without a carrier, may be applied to packaging materials. More specifically, the benzoxaborole treatments of the present disclosure may be applied, imbedded, impregnated, or coated onto or into one or more surfaces of a packaging material (e.g., PET clamshells or liner materials) or a device (e.g., a container or a chamber), collectively and interchangeably called a "chamber." When coated onto or imbedded into surfaces of packaging materials, the benzoxaborole compound treatment volatilizes to treat food products, which ultimately preserves freshness of the food. Thus, the compound treatments of the present disclosure allow for the uniform treatment of food packaging materials to protect food products comprised therein.

A chamber of the present disclosure may be any container in which a food product may be comprised therein for harvest, storage, and/or retail usage. For example, a chamber may be made of any material sufficient to hold food including, but not limited to, cardboard, paper, paperboard, corrugated paper, plastic (e.g., thermosets and thermoplastics), glass, polystyrene, cellulosic material, metals (e.g., aluminum, foils, laminates, tinplate, and/or steel, such as tin- free steel), or any other semipermeable or impermeable material. Exemplary chambers of the present disclosure may be made of polyester, such as polycarbonate, polyethylene naphthalate, and polyethylene terephthalate (i.e., PET or PETE). Thus, an illustrative embodiment of a chamber of the present disclosure is a PET clamshell.

The chamber of the present disclosure may be of any size to hold food products within the packaging materials, such as an individual or singulated chamber embodiment. For example, illustrative individual chambers may have a volume ranging from about 0.1 liters (L) to about 50 L, from about 0.1 L to about 40 L, from about 0.1 L to about 30 L, from about 0.1 L to about 20 L, from about 0.1 L to about 10 L, from about 0.1 L to about 5 L, from about 0.1 L to about 4 L, from about 0.1 L to about 3 L, from about 0.1 L to about 2 L, from about 1 L to about 50 L, from about 5 L to about 40 L, from about 20 L to about 40 L, from about 25 L to about 50 L, from about 30 L to about 40 L, from about 35 L to about 40 L, and at about 0.1 L, about 0.2 L, about 0.3 L, about 0.4 L, about 0.5 L, about 1 L, about 2 L, about 10 L, about 20 L, about 30 L, about 35 L, about 40 L, and about 50 L.

An additional chamber embodiment may be capable of holding a plurality of individual chambers. A plurality of individual chambers, such a PET clamshells, may include two or more to thousands, to many thousands to tens or hundreds of thousands or millions of PET

clamshells. For example, one box may comprise about 384 clamshells, and a chamber embodiment may comprise thousands of boxes of clamshells (e.g., from about 384,000 to about 3,840,000 to about 384,000,000 of clamshells). Thus, a plurality of PET clamshells is also an illustrative embodiment of the chamber of the present disclosure, which is particularly utilized for large-scale and/or commercial treatment methods of chambers.

A further embodiment of the chamber of the present disclosure may comprise a liquid- absorbing material. The liquid- absorbing material may be comprised within the chamber, such as on the internal top, bottom or side panels of the chamber. The liquid-absorbing material may be comprised on the exterior of the chamber, such as on the external top, bottom or side panels of the chamber. The liquid- absorbing material may also be comprised on or in one or more liners, wrapping, labels, tags, stickers, pads, or other packing components located, attached, and/or affixed to the inside or the outside of the chamber.

The liquid-absorbing material may comprise any material that is able to absorb and retain a liquid composition of the active compound. For example, illustrative embodiments of the liquid- absorbing material include, but are not limited to, cotton, paper, foam, etc.

Absorption of an active ingredient (i.e., benzoxaborole) into the liquid-absorbing material enables the liquid-absorbing material to serve as a reservoir capable of releasing the benzoxaborole treatment to the food product comprised in the chamber over a time period. The liquid-absorbing material may provide for slow-release or quick-release of the benzoxaborole treatment to the food product. Thus, the liquid-absorbing material enables differential treatment of the food product based on the time period required for protection of food products comprised therein. For example, food products that need limited antimicrobial protection may be packaged in a chamber comprising a quick-release liquid-absorbing material, while food products requiring an extended term of antimicrobial protection may be packaged in a chamber comprising a slow-release liquid- absorbing material.

Slow-release liquid- absorbing materials include, but are not limited to, materials that enable the release of the active ingredient to the food product for a time period of over 12 hours, such as from over 12 hours to about 31 days, including from over 12 hours to about 25 days, from over 12 hours to about 20 days, from over 12 hours to about 15 days, from over 12 hours to about 10 days, from over 12 hours to about 5 days, from over 12 hours to about 30 days, from over 12 hours to about 24 days, from about 24 hours to about 30 days, from about 2 days to about 28 days, from about 3 days to about 25 days, from about 4 days to about 20 days, and about 5 days, about 10 days, about 15 days, about 20 days, about 25 days, about 30 days, and any number of days between 1 day to 30 days.

Quick-release liquid-absorbing materials include, but are not limited to, materials that enable the release of the active ingredient to the food product for a time period ranging from about 12 hours or less, such as from about 5 seconds to about 12 hours, from about 5 seconds to about 10 hours, from about 10 seconds to about 8 hours, from about 15 seconds to about 6 hours, from about 20 seconds to about 4 hours, from about 25 seconds to about 2 hours, from about 5 seconds to about 1 hour, from about 10 seconds to about 45 minutes, from about 15 seconds to about 30 minutes, from about 20 seconds to about 15 minutes, from about 25 seconds to about 5 minutes, from about 5 seconds to about 1 minute, from about 5 seconds to about 30 seconds, from about 5 seconds to about 15 seconds, and from about 5 seconds to about 10 seconds.

The chamber may also comprise one or more holes or apertures. The apertures may have any shape, and may have a size ranging from about 2 mm to about 2 cm, and from about 2.5 mm to about 1.5 cm, from about 5 mm to about 1.5 cm, from about 7.5 mm to about 1.25 cm, from about 10 mm to about 1 cm, from about 15 mm to about 0.75 cm, from about 20 mm to about 0.5 cm, and from about 25 mm to about 0.25 cm. In addition, the apertures may be in any location on the chamber material, but typically, the apertures are located at the base, the lid, the sides, or a combination thereof on the chamber. The apertures allow for introduction of treatment to the chamber and/or release of treatment from the chamber.

Upon introduction of compound treatment into or onto the chamber, the apertures permit uniform distribution of treatment vapor, gas, or fog particles throughout the chamber. The apertures also allow for full drainage, venting, and/or release of the unused portion of the treatment or treatment carrier from the chamber. Unused treatment and/or treatment carrier may be recycled to treat subsequent and/or additional containers, materials, or chambers.

An illustrative example of a product of the method described herein is one or more benzoxaborole-treated PET clamshell, such as a plurality of PET clamshells. PET clamshells are commonly used to transport strawberries and other soft fruits. Therefore, a benzoxaborole- treated PET clamshell would provide the greatest protection to the fruit contained therein since the active ingredient is coated on the surfaces of the primary packaging of the fruit. Primary protection of the fruit could also occur via treatment of a material contained within the PET clamshell, such as a liquid- absorbing material in the form of a tag, a pad, or other embodiments described herein. Secondary protection of the fruit would occur by applying the active ingredient to a liner, a box, a bag, a wrap or other packaging material in which the primary chambers are placed for storage or transport.

METHODS OF ADMINISTERING BENZOXABOROLE COMPOUNDS

The present disclosure is directed to methods of uniformly treating food products by providing antimicrobial protection to food, such as plants, crops, and meats. The present methods are directed to large-scale treatment of food packaging materials to uniformly protect plants from plant pathogens and microorganisms that cause food decay. More specifically, plant pathogens that inhibit, reduce, or compromise food freshness may be treated, prevented, or eradicated by the methods described herein.

Exemplary, microorganisms encompassed by the present disclosure include, but are not limited to, Botrytis cinerea, Mucor piriformis, Fusarium sambucinum, Aspergillus brasiliensis, and Peniciliium expansum. Additional pathogens encompassed by the present invention include, but are not limited to Acremonium spp., Albugo spp., Alternaria spp., Ascochyta spp., Aspergillus spp., Botryodiplodia spp., Botryospheria spp., Botrytis spp., Byssochlamys spp., Candida spp., Cephalosporium spp., Ceratocystis spp., Cercospora spp., Chalara spp., Cladosporium spp., Colletotrichum spp., Cryptosporiopsis spp., Cylindrocarpon spp.,

Debaryomyces spp., Diaporthe spp., Didymella spp., Diplodia spp., Dothiorella spp., Elsinoe spp., Fusarium spp., Geotrichum spp., Gloeosporium spp., Glomerella spp., Helminthosporium spp., Khuskia spp., Lasiodiplodia spp., Macrophoma spp., Macrophomina spp., Microdochium spp., Monilinia spp., Monilochaethes spp., Mucor spp., Mycocentrospora spp., Mycosphaerella spp., Nectria spp., Neofabraea spp., Nigrospora spp., Peniciliium spp., Peronophythora spp., Peronospora spp., Pestalotiopsis spp., Pezicula spp., Phacidiopycnis spp., Phoma spp., Phomopsis spp., Phyllosticta spp., Phytophthora spp., Polyscytalum spp., Pseudocercospora spp., Pyricularia spp., Pythium spp., Rhizoctonia spp., Rhizopus spp., Sclerotium spp., Sclerotinia spp., Septoria spp., Sphaceloma spp., Sphaeropsis spp., Stemphyllium spp., Stilbella spp., Thielaviopsis spp., Thyronectria spp., Trachysphaera spp., Uromyces spp., Ustilago spp., Venturia spp., and Verticillium spp., and bacterial pathogens, such as Bacillus spp.,

Campylobacter spp., Clavibacter spp., Clostridium spp., Erwinia spp., Escherichia spp., Lactobacillus spp., Leuconostoc spp., Listeria spp., Pantoea spp., Pectobacterium spp.,

Pseudomonas spp., Ralstonia spp., Salmonella spp., Shigella spp., Staphylococcus spp., Vibrio spp., Xanthomonas spp., and Yersinia spp.

Benzoxaborole treatments may be applied, administered, or coated on the inside or the outside of a chamber or packaging material. When the benzoxaborole treatment is in any form, but particularly in liquid, spray, vapor, or gas form, a drying step may be provided in the present method that allows excess treatment carrier to dry. This step may also produce a residue of the active benzoxaborole ingredient at the proper levels of efficacy on the surface of the food packaging material in order to provide extended antimicrobial control and inhibition for plants, crops, and meats contained therein.

Any food product, including plants, crops, or meats, may be treated using the present method. Minimally-processed packaged products (e.g., packaged vegetables, fruits, or meats) may also be treated with the method described herein. Horticultural crops of the present method include, but are not limited to, vegetable crops, fruit crops, edible nuts, flowers and ornamental crops, nursery crops, aromatic crops, and medicinal crops.

Plants and agricultural crops in any production cycle may be used in the method of the present application. For example, post-harvest plants and crops may be treated during field packing, palletization, in-box, storage, and throughout the distribution network. Further, plants being transported by any mode, including, but not limited to local vehicles, transport trailers, marine containers, aircraft containers, etc. may be treated using the method described herein.

Ideally, the chamber or plurality of chambers of the present disclosure are treated prior to use during food field-packaging of plants, meats, or crops, such as soft fruits. For example, treated chambers may be located, stored, and/or kept at the site of clamshell manufacturers, at the central facility of farmers or ranchers, or in a portable unit for immediate in-field treatments. Additionally, treated chambers or materials may be provided to a food producer directly from the manufacturer, wherein the manufacturer has previously applied benzoxaborole to the surface of the packaging. Alternatively, a food producer may independently use a method, machine, or instrument to treat containers or packaging materials with the benzoxaborole compositions as described herein.

Large-scale treatment of food product packaging materials and chambers are comprised in the methods of the present disclosure. Large-scale treatment comprises treatment of chambers or a plurality of chambers in mass, and typically for commercial and/or industrial use. For example, the methods of the present disclosure may comprise treating a plurality of chambers with the benzoxaborole active ingredient described herein. Large-scale treatment methods of the present disclosure may occur before the chamber has been formed (i.e., preformation), during formation of the chamber (i.e., formation), and after the chamber has been formed (i.e., postformation). Formation is the process of forming or producing one or more chambers of the present disclosure, which may include thermoforming.

Preformation treatment of the chamber comprises contacting a packaging material, such as plastic, that will be formed into the chamber with an active ingredient of the present disclosure (e.g., benzoxaborole) prior to the beginning of the formation process. Formation comprises treatment of the packaging material of the chamber with the active ingredient after the formation process has started. Postformation treatment of the chamber occurs when the packaging material has been formed into a chamber and the chamber is then treated with the active ingredient. For example, the chambers may be treated with the active ingredient prior to, during, or after formation of the chamber using methods including, but not limited to, dipping, drenching, spraying, painting, vaporizing, and/or sublimation.

One embodiment of the large-scale method described herein comprises use of a printer to print the active compound treatment on the lining or material of a plurality of chambers. Another embodiment of the large-scale treatment method comprises spraying the plurality of chambers with the active compound treatment. A further embodiment of the large-scale treatment method comprises dipping a plurality of chambers into a vat of active compound treatment and removing the treated chambers to dry onto the chamber material. Additionally, fogging or spraying a plurality of chambers with the active compound during manufacturing in an industrial-sized device is also an embodiment of the large-scale method of the present disclosure.

Food products may or may not be inside of the chamber during application of the benzoxaborole treatment. If the food product is already inside the chamber, treatment of the chamber with the active ingredient may be applied while the chamber is open, closed, or sealed. Typically, however, after the treated chambers are produced, food products, such as plants, crops, or meats, may be manually or robotically (e.g., by a machine) placed in the treated chamber in preparation for antimicrobial treatment of the food.

The proximity and/or distance between the emitting source of active ingredient and the food product is critical. Notably, the distance from the benzoxaborole coated surfaces of the packaging material or chamber and the food product is inversely related to the efficacy of antimicrobial protection of the food. In other words, the greater the distance between the coated/treated surfaces of the material or chamber and the food product, the lesser is the level of antimicrobial protection conveyed to the treated food product, including plants, crops, or meat.

Related to this property, treated surfaces of a chamber or packaging material, such as a bag, a box, a wrap, a liner, or other packaging material that is placed over an entire pallet of clamshells of food products may be less effective in delivering the active ingredient to the food than a treated surface that is immediately adjacent to or in contact with the plant, crop, or meat food products, such as the clamshell surfaces themselves. Therefore, having the treatment coated directly on a surface or imbedded into the individual product chamber that is the primary packaging material, meaning the first layer of packaging of the food product, provides the greatest antimicrobial protection to the food product. Similarly, treating the internal surface of the primary packaging material, such as a chamber, provides even greater antimicrobial effect than treating an external surface of the chamber. Therefore, the distance between the emitting source of the active ingredient and the food product should be minimized for best results.

More specifically, the distance between the emitting source and the food product should remain less than about 6 feet. In an illustrative embodiment, the distance between the emitting source and the food product ranges from about 0.1 inches to about 6 feet, from about 0.5 inches to about 5 feet, from about 1 inch to about 4 feet, from about 1.5 inches to about 3 feet, from about 2 inches to about 2 feet, from about 0.5 inches to about 12 inches, from about 1 inch to about 24 inches, from about 0.5 inches to about 6 inches, from about 0.5 inches to about 5 inches, from about 0.5 inches to about 4 inches, from about 0.5 inches to about 3 inches, from about 0.5 inches to about 2 inches, and from about 0.5 inches to about 1 inch. Close proximity of the emitting source of the benzoxaborole active ingredient applied to the primary PET clamshell ensures that the food product is exposed to the active ingredient which inhibits microorganisms that may infect the food.

An exemplary embodiment of the method described herein comprises vaporizing or subliming the benzoxaborole compound or molecule into a gaseous form. The benzoxaborole compound gas or vapor may be at any concentration that allows the compound or molecule to adhere to the chamber (e.g., a clamshell) or packaging surfaces prior to taking the chamber or material into a field for the harvest operation.

For example, the benzoxaborole compound vapor or gas may be effectively

administered to a chamber at a concentration ranging from about 0.1 mg/chamber to about 10 mg/chamber, from about 0.1 mg/chamber to about 8 mg/chamber, from about 0.1 mg/chamber to about 7 mg/chamber, from about 0.1 mg/chamber to about 6 mg/chamber, from about 0.1 mg/chamber to about 5.5 mg/chamber, from about 0.1 mg/chamber to about 5 mg/chamber, from about 0.1 mg/chamber to about 4 mg/chamber, from about 0.1 mg/chamber to about 3.5 mg/chamber, from about 0.1 mg/chamber to about 3.2 mg/chamber, from about 0.1 mg/chamber to about 2 mg/chamber, from about 0.1 mg/chamber to about 1.5 mg/chamber, from about 0.1 mg/chamber to about 1 mg/chamber, from about 0.1 mg/chamber to about 0.35 mg/chamber, from about 0.1 mg/chamber to about 0.32 mg/chamber, from about 0.1 mg/chamber to about 0.25 mg/chamber, from about 0.1 mg/chamber to about 0.22 mg/chamber, 0.2 mg/chamber to about 8 mg/chamber, from about 0.2 mg/chamber to about 7 mg/chamber, from about 0.2 mg/chamber to about 6 mg/chamber, from about 0.2 mg/chamber to about 5.5 mg/chamber, from about 0.2 mg/chamber to about 5 mg/chamber, from about 0.2 mg/chamber to about 4 mg/chamber, from about 0.2 mg/chamber to about 3.5 mg/chamber, from about 0.2 mg/chamber to about 3.2 mg/chamber, from about 0.2 mg/chamber to about 2 mg/chamber, from about 0.2 mg/chamber to about 1.5 mg/chamber, from about 0.2 mg/chamber to about 1 mg/chamber, from about 0.2 mg/chamber to about 0.35 mg/chamber, from about 0.2

mg/chamber to about 0.32 mg/chamber, from about 0.2 mg/chamber to about 0.25 mg/chamber, from about 0.2 mg/chamber to about 0.22 mg/chamber, 0.2 mg/chamber to about 8

mg/chamber, from about 0.3 mg/chamber to about 7 mg/chamber, from about 0.3 mg/chamber to about 6 mg/chamber, from about 0.3 mg/chamber to about 5.5 mg/chamber, from about 0.3 mg/chamber to about 5 mg/chamber, from about 0.3 mg/chamber to about 4 mg/chamber, from about 0.3 mg/chamber to about 3.5 mg/chamber, from about 0.3 mg/chamber to about 3.2 mg/chamber, from about 0.3 mg/chamber to about 2 mg/chamber, from about 0.3 mg/chamber to about 1.5 mg/chamber, from about 0.3 mg/chamber to about 1 mg/chamber, from about 0.3 mg/chamber to about 0.35 mg/chamber, from about 0.3 mg/chamber to about 0.32 mg/chamber, and at about 0.2 mg/chamber, 0.316 mg/chamber, 1 mg/chamber, 3.16 mg/chamber, and 5 mg/chamber.

Alternatively, the benzoxaborole compound may be prepared as a liquid formulation. Preparing a liquid composition of the benzoxaborole treatment requires mixing the

benzoxaborole compound with a liquid carrier, such as a solvent or water or combination thereof. Once the liquid treatment is prepared, the chamber or material is drenched or flooded with the liquid treatment, such that the liquid treatment touches all internal surfaces. Most of the liquid treatment carrier may then be drained from an aperture or hole in the chamber or material. After the carrier is substantially removed from the chamber, residue of the active ingredient (i.e., benzoxaborole) in the remaining liquid is allowed to dry, such as at room temperature (e.g., about 21°C to about 23°C).

Drying of the treatment composition, including the liquid carrier, may occur

instantaneously or within seconds (sees). In particular, the higher the concentration of active ingredient (i.e., benzoxaborole) and the lower the volume of compound treatment, the less time is required for drying the treatment composition onto the chamber. For example, drying time of the treatment composition onto the chamber material may range from about 0.1 sees to about 60 sees, from about 0.2 sees to about 45 sees, from about 0.3 sees to about 30 sees, from about 0.4 sees to about 20 sec, from about 0.5 sees to about 15 sees, from about 1 sec to about 10 sees, from about 5 sees to about 60 sees, and at about 5 sees. Once dried, the active benzoxaborole ingredient is coated onto the surface of the chamber to provide immediate antimicrobial protection to food, such as plants, crops, or meats placed therein.

As previously mentioned, a liquid benzoxaborole treatment may be administered to a fruit or a vegetable clamshell, such that all internal surfaces of the chamber come into contact with the active ingredient. After draining the treatment carrier (e.g., water or solvent) from the clamshell, warm air (i.e., room temperature) is applied to dry the remaining liquid on the clamshells. The treated clamshells comprising the coated surfaces and active ingredient may then go immediately to the field for picking operations. Food, such as fresh fruits and berries placed within the treated clamshells, is protected from disease microorganisms by the volatile active ingredient (i.e., benzoxaborole) emitted from the surface of the clamshells from the time the fruit is placed inside of the chamber.

In yet another embodiment of the present method, the benzoxaborole active ingredient may be administered by fogging as a fine mist into a suitable chamber. The compound may be fogged using any cold, thermal, ultrasonic, or similar fogging-based technologies. The micron particles of water, solvent, or other carriers in the formulations may assist distribution and deposition of the benzoxaborole particles on the chamber surfaces. Upon drying, the method will result in a thin coating of active ingredient on the chamber surfaces. This thin coating of benzoxaborole will volatilize over time, and uniformly protect the food contents of the chamber from pathogenic infection and decay.

Accordingly, the method described herein provides a large-scale administration of an antimicrobial agent, such as benzoxaborole, to the packaging surfaces of food product chambers. Importantly, the presently described method enables greater uniformity and consistency of application of the active benzoxaborole treatment composition to the food packaging materials. Ultimately, the present large-scale method of uniformly treating food chambers results in a significantly extended time period (i.e., up to about 31 days or a full month) of antimicrobial protection of the food product comprised within the treated chambers.

EXAMPLES

Illustrative embodiments of the methods of the present disclosure are provided herein by way of examples. While the concepts and technology of the present disclosure are susceptible to broad application, various modifications, and alternative forms, specific embodiments will be described here in detail. It should be understood, however, that there is no intent to limit the concepts of the present disclosure to the particular forms disclosed, but on the contrary, the intention is to cover all modifications, equivalents, and alternatives consistent with the present disclosure and the appended claims.

The following experiments were used to determine the effect of different concentrations of benzoxaborole compounds when administered onto the surfaces of packaging materials or chambers by various application techniques. In the following experiments, benzoxaborole treatment compositions are applied to chambers such that food products may be exposed to the antimicrobial treatment for an initial time period. For example, food products may be exposed to the benzoxaborole treatment on the surfaces of the treated chamber for the initial time period ranging from less than 1 day to about 8 days, and at about 5 days. Treatment temperatures of the chamber during the initial time period ranged from about 0.5°C to about 5°C, and at about 1°C.

After the initial time period in which the food is exposed to the active ingredient of the treated chamber, the chamber may be unsealed (if previously sealed), and allowed to vent for a secondary time period. The food may remain in the chamber for the secondary time period ranging from about 1 day to about 8 days, and at about 6 days. The temperature of the chamber during the second time period remains at room temperature, which ranges from about 20°C to about 23°C, and at about 21°C.

After expiration of the secondary time period, inhibition of plant pathogens and infection may be assessed. For example, in vitro samples may have the growth of the microorganism or pathogen on agar or in media assessed, evaluated, and compared to a control sample where no benzoxaborole treatment was administered. Similarly, in vivo samples may have the severity and incidence of pathogenic disease assessed, evaluated, and compared to a control sample where no benzoxaborole treatment was administered or different treatment conditions were applied. Example 1: Benzoxaborole Compound Treatment of Fruit Clamshells by

Spraying, Painting, and Sublimation {In Vivo)

An in vivo assay was used to evaluate the ability of Compound A to volatilize from a clamshell chamber and control pathogenic infection when applied by various techniques.

Multiple empty 1-lb PET clamshells (ProducePackaging.com, #036QT) were placed inside triplicate air-tight 36 L chamber (Fisher Scientific, Catalogue #08-642-23C) fitted with a bulkhead septum port (Swagelok, SS-401-61, Solon, OH).

An appropriate amount of Compound A, to achieve a final treatment rate of 5 mg per clamshell (i.e., 5 mg/clamshell), was dissolved in acetone and 100 μΐ ^ of the solution was pipetted into a small glass tube. The tube was then placed inside a pre-heated sublimation device (0.5" OD by 6" long thermostatically healed copper tube mounted to a 2 L/min aquarium pump) set at 60°C for 1 minute to allow the acetone to evaporate. Compound A was then introduced into the cabinets of the chamber through the bulkhead port containing the clamshells by using the sublimation device set at 180°C. Compound A headspace was permitted to equilibrate overnight at 21°C

Five milligrams of Compound A was dissolved in 1 ml of ethanol prior to being uniformly administered to the interior of the clamshell by painting or spraying, and then dried for 5 minutes. After coating the clamshells with Compound A using various application techniques (i.e., sublimation, spraying, or painting), eight ethanol-washed strawberries were placed in the clamshell with stem end facing downwards. Each strawberry fruit was wounded using a T15 screwdriver tip to a uniform depth of eight mm (8mm). Each fruit wound was inoculated with 20 μL· of lxlO 5 spores/ml pathogen spore suspension of Botrytis cinerea, which is a fungal pathogen known to cause gray mold infection of fruits, such as grapes and strawberries. Uninoculated strawberries were removed from their commercial package, and directly transferred into treated clamshells without any washing or inoculation steps.

Treated clamshells lids were closed, and then placed at 1°C for an initial time period of 5 days. Clamshells were then removed from low temperature, and held for a second time period of 6 days at room temperature where the point of inoculation on the strawberry fruits was assessed for indication of disease incidence reported as a percentage (%). Severity of disease incidence was also reported. Disease severity was rated on a scale ranging from 0 to 4, where "0" indicated no disease severity, "1" indicated minimal disease severity, "2" indicated medium disease severity, "3" indicated high disease severity, and "4" indicated exceptionally high disease severity.

The outcome of this in vivo experiment is summarized in Table 1. Results demonstrate good in vivo antimicrobial activity of Compound A against B. cinerea, with a reduction in disease incidence and severity with all three application techniques (i.e., painting, spraying, and sublimation). In particular, each method of treating clamshells showed significant inhibition of gray mold incidence and severity in strawberries as compared to control. More specifically, on Days 1-6, the percentage of gray mold incidence increased from 30.5% to 100% and 0% to a maximum of 0.9% in control fruits and treated inoculated fruits, respectively. Even in uninoculated fruit, the percentage of gray mold incidence increased from 1.5% to 100% and 0% to a maximum of 21.7% in control fruits and treated fruits, respectively. In both inoculated and uninoculated fruits, the spraying technique was comparable to or better than painting or sublimation in minimizing the incidence or severity of gray mold. Ultimately, treating the clamshells with benzoxaborole Compound A significantly inhibited the growth of B. cinerea in the strawberries and preserved the freshness of the fruit for at least 3 days longer than the untreated strawberries.

Example 2: Benzoxaborole Compound Treatment of Clamshells Containing Agar Plates by

Spraying, Painting, and Sublimation (In Vitro)

An in vitro assay was used to evaluate the ability of Compound A to volatilize from a clamshell to control fungal pathogenic infection when applied to the clamshell by various application techniques. Multiple empty 1-lb PET clamshells (ProducePackaging.com,

#036QT) were placed inside triplicate air-tight 36 L chamber (Fisher Scientific, Catalogue #08- 642-23C) fitted with a bulkhead septum port (Swagelok, SS-401-61, Solon, OH).

An appropriate amount of Compound A to achieve a final treatment rate of 5 mg/clamshell, 1 mg/clamshell, or 0.2 mg/clamshell, was dissolved in acetone and 100 μΐ ^ of the solution was pipetted into a small glass tube. The tube was then placed inside a pre -heated sublimation device (0.5" OD by 6" long thermostatically healed copper tube mounted to a 2 L/min aquarium pump) set at 60°C for 1 minute to allow the acetone to evaporate. Compound A was then introduced into the cabinets of the chamber through the bulkhead port containing the clamshells by using the sublimation device set at 180°C. Compound A headspace was permitted to equilibrate overnight at 21°C.

For paint and spray applications, 5 mg of Compound A was dissolved in 1 ml of ethanol prior to uniformly painting or spraying the interior of the clamshell. After spraying or painting, the clamshell was then permitted to dry for 5 minutes. After coating the clamshells with Compound A using various applications (i.e., sublimation, spraying, or painting), 10-cm Petri plates containing half strength Potato Dextrose Agar were inoculated with 1 of 1 x 10 5 spores/ml Botrytis cinerea spore suspension. The inoculated petri plates were then sealed with a breathable film (AeraSeal; P/N: B- 100, Excel Scientific, Victorville, CA), and placed inside the treated clamshell.

To determine the period of time coated packaging could release effective levels of the active ingredient, treated clamshells containing the inoculated plates were then placed inside a 2.55 L SnapWare airtight container (Model# 109842) for three (3) days at 21°C (Series I). After incubation, plates were removed and cultures were evaluated for percent growth relative to a control based on measurement of fungal colony diameter (mm).

Table 1. Comparison of in-clamshell Compound A application techniques to control growth of Botrytis cinerea inoculated strawberries as compared to uninoculated fruit.

Gray Mold Incidence (%)

Inoculated Fruit Day O Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

Control 30.5 62.1 84.3 93.5 100.0 100.0 100.0

Paint 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Spray 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Sublimation 0.0 0.0 0.0 0.0 0.0 0.9 0.9

Gray Mold Severity (0-4)

Day O Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

Control 0.2 0.4 1.1 2.1 2.6 4.0 4.0

Paint 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Spray 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Sublimation 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Gray Mold Incidence (%)

Uninoculated

Day O Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

Fruit

Control 1.5 7.1 39.9 96.7 96.7 100.0 100.0

Paint 0.0 0.3 3.9 5.1 7.4 17.0 21.7

Spray 0.0 0.6 2.1 4.8 6.0 17.9 19.9

Sublimation 0.0 0.3 1.5 3.3 5.7 16.7 20.2

Gray Mold Severity (0-4)

Day O Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

Control 0.3 0.9 1.5 2.8 3.5 4.0 4.0

Paint 0.0 0.1 0.5 0.8 0.9 1.1 1.1

Spray 0.0 0.2 0.3 0.5 0.7 1.2 1.3

Sublimation 0.0 0.1 0.3 0.7 0.9 1.1 1.3

Meanwhile, freshly prepared inoculated plates were placed inside the clamshell for an additional three (3) days of incubation (Series 2). This process, where freshly prepared inoculated plates were placed inside the clamshell, was repeated twice more, to produce a 12 full day evaluation of 4 series of 3 days each. The outcome of this in vitro experiment is summarized in Table 2. Results demonstrate good volatile in vitro antimicrobial activity of Compound A against Botrytis cinerea with all three application techniques (i.e., painting, spraying, and sublimation). In addition, greater inhibition of pathogenic growth was observed at higher treatment rates.

In particular, each technique of treating clamshells showed significant inhibition of mycelial growth. More specifically, administration of 5 mg/clamshell of benzoxaborole treatment by all three techniques were effective to completely inhibit mycelial cell growth the first six days (Table 2). In the remaining six days, sublimation most effectively inhibited mycelial growth (84.4%), followed by spraying (64.9%), and painting (44.7%). At lower treatment concentrations (i.e., 1 mg/clamshell and 0.2 mg/clamshell), all of the treatment techniques failed to inhibit mycelial cell growth by Days 6 to 9. Ultimately, treating the clamshells with benzoxaborole Compound A significantly inhibited the growth of B. cinerea inoculated on agar plates placed therein for time periods typical of fruit storage by the supply chain and consumers.

Table 2. Comparison of treatment techniques of Compound A to volatilize from clamshells and provide in vitro inhibition of Botrytis cinerea

Rate Series 1 Series 2 Series 3 Series 4

Method (mg/clamshell) (0 to 3 days) (3 to 6 days) (6 to 9 days) (9 to 12 days)

Sublimation 5 100.0 100.0 83.1 84.4

1 90.1 0.0 0.0

0.2 90.9 0.0 0.0

Paint 5 100.0 100.0 57.5 44.7

1 100.0 18.8 0.0 —

0.2 3.4 0,0 0.0 —

Spray 5 100.0 100.0 61.2 64.9

1 100.0 18.6 0.0

0.2 81.2 0.0 0.0 Example 3: Dose Response of Benzoxaborole Compound Treatment of Fruit

Clamshells by Vapor-Coating {In Vivo)

This in vivo assay was used to evaluate the ability of vaporized Compound A to volatilize from a clamshell in order to control or inhibit pathogenic microorganisms. This experiment was conducted exactly as described in Example 1, with a few exceptions. After equilibrating the clamshell overnight at 21°C, Compound A was administered to the clamshell as a vapor only. In addition, Compound A was vapor-coated onto the surface of the clamshell at a final treatment rate of 3.16 mg/clamshell, 1 mg/clamshell, or 0.316 mg/clamshell. After vapor-coating application, Compound A headspace was permitted to equilibrate overnight at 21°C. Strawberries were inoculated with Botrytis cinerea, placed within the clamshells for an initial time period, and assessed for disease incidence and severity over a second time period as described in Example 1.

The outcome of this in vivo experiment is summarized in Table 3. Results demonstrate good in vivo antimicrobial activity of Compound A against B. cinerea, with a greater reduction in disease incidence and severity observed at higher treatment rates.

In particular, each concentration of active ingredient on treated clamshells showed inhibition of gray mold severity in inoculated strawberries as compared to control (see Table 3). More specifically, on Days 1-6, the percentage of gray mold severity increased from 0.8 to 4.0 in control fruits as compared to 0 to 0.2 and 0 to 0.7 for inoculated fruits treated with 3.16 mg/clamshell or 1 mg/clamshell of Compound A, respectively. At the lower treatment concentration of 0.316 mg/clamshell, the percentage of gray mold severity increased from 0.4 to 3.9. Ultimately, treating the clamshells with different concentrations of benzoxaborole Compound A significantly inhibited the growth of B. cinerea in the strawberries in a dose dependent manner.

Table 3. Dose response of in-clamshell Compound A applied by sublimation to control growth of

Botrytis cinerea inoculated on strawberries.

Inoculated Fruit Gray Mold Severity (0-4)

Rate (mg per clamshell) Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

3.16 0.0 0.0 0.0 0.0 0.1 0.2

1.00 0.0 0.0 0.0 0.1 0.4 0.7

0.316 0.4 1.3 2.2 3.4 3.8 3.9

Control 0.8 1.8 2.7 3.6 4.0 4.0 Example 4: Dose Response of Benzoxaborole Compound Treatment of Fruit

Clamshells by Spraying {In Vivo)

This in vivo assay was used to evaluate the ability of different concentrations of

Compound A to volatilize from clamshell in order to control or inhibit fruit infection by pathogenic microorganism, Botrytis cinerea. This experiment was conducted exactly as described in Example 1, with a few exceptions. After equilibrating the clamshell overnight at 21°C, Compound A was administered to the clamshells by spraying only. Compound A was sprayed onto the surface of the clamshells at a final treatment rate of 5 mg/clamshell or 1 mg/clamshell. The clamshells were then permitted to dry for 5 minutes. Strawberries were inoculated with Botrytis cinerea, placed within the clamshells for the initial time period (i.e., 5 days), and assessed for disease incidence and severity over the second time period (i.e., 6 days) as described in Example 1.

The outcome of this in vivo experiment is summarized in Table 4. Results demonstrate good in vivo antimicrobial activity of Compound A against B. cinerea, with a reduction in disease incidence and severity with both 5 mg/clamshell and 1 mg/clamshell concentrations applied by spraying. In particular, each concentration of treating clamshells showed inhibition of gray mold incidence and severity in strawberries as compared to control. On Days 1-6, the percentage of gray mold incidence increased from 6.3% to 100% in control inoculated fruits, while there was no growth in 5 mg/clamshell-treated inoculated fruits. Even the 1

mg/clamshell-treated inoculated fruits inhibited gray mold incidence to a 52.5% maximum. In uninoculated fruit, the percentage of gray mold incidence increased from 0.5% to 78.5% in control fruits, and similarly, from 0% to 100% in 1 mg/clamshell-treated uninoculated fruits. However, the percentage of gray mold incidence only increased from 0.5% to 78.5% in 5 mg/clamshell-treated uninoculated fruits.

In addition, each concentration of active ingredient on treated clamshells showed significant inhibition of gray mold severity in inoculated strawberries as compared to control (see Table 4). More specifically, on Days 1-6, the degree of gray mold severity increased from 0.0 to 4.0 in control fruits as compared to 0 to 1.5 in 1 mg/clamshell-treated inoculated fruits and no growth in 5 mg/clamshell-treated inoculated fruits. For uninoculated fruits, both control and 1 mg/clamshell-treated fruits showed a gray mold severity level of 4.0 by Day 4, while the 5 mg/clamshell-treated uninoculated fruits only showed a gray mold severity level of 3.5 on Day 6. Ultimately, these data demonstrate that treating clamshells with different concentrations of benzoxaborole Compound A significantly inhibited the gray mold infection of B. cinerea inoculated in strawberries in a dose dependent manner.

Table 4. Dose Response of in-clamshell Compound A application techniques to control growth of Botrytis cinerea inoculated in strawberries as compared to uninoculated strawberries.

Inoculated Fruit Gray Mold Incidence ( %)

Rate (mg/clamshell) Day O Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

5 0.0 0.0 0.0 0.0 0.0 0.0 0.0

1 0.0 0.0 1.3 3.8 11.3 36.3 52.5

Control 6.3 72.5 100.0 100.0 100.0 100.0 100.0

Gray Mold Severity (0-4)

Day O Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

5 0.0 0.0 0.0 0.0 0.0 0.0 0.0

1 0.0 0.0 0.0 0.1 0.2 0.6 1.5

Control 0.0 0.7 1.4 2.7 3.6 3.8 4.0

Uninoculated Fruit Gray Mold Incidence (%)

Rate (mg/clamshell) Day O Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

5 0.5 0.5 0.9 3.3 16.2 31.9 78.5

1 0.0 1.9 21.6 89.5 100.0 100.0 100.0

Control 1.0 6.0 46.0 100.0 100.0 100.0 100.0

Gray Mold Severity (0-4)

Day O Day 1 Day 2 Day 3 Day 4 Day 5 Day 6

5 0.1 0.1 0.3 0.5 1.0 1.4 3.5

1 0.0 0.5 1.8 3.6 4.0 4.0 4.0

Control 0.3 1.0 2.9 4.0 4.0 4.0 4.0

Example 5: Benzoxaborole Compound Treatment of Fruit Clamshells by

Sublimation {In Vivo)

This in vivo assay was used to evaluate the ability of Compound A to volatilize from a clamshell in order to control or inhibit fruit infection by pathogenic microorganism, Botrytis cinerea. This experiment was conducted exactly as described in Example 1, with a few exceptions. After equilibrating the clamshell overnight at 21°C, Compound A was

administered to the clamshell by sublimation only. Compound A was sublimed onto the surface of the clamshell at a final treatment rate of 5 mg/clamshell. Strawberries were inoculated with Botrytis cinerea, placed within the clamshell for an initial time period of 6 days, and assessed for disease incidence and severity over a second time period of 7 days as described in Example 1.

The outcome of this in vivo experiment is summarized in Table 5. Results demonstrate good in vivo antimicrobial activity of Compound A against B. cinerea, with a reduction in disease incidence and severity with the 5 mg/clamshell concentrations applied by sublimation. In particular, both inoculated and uninoculated 5 mg/clamshell-treated clamshells showed inhibition of gray mold incidence and severity in strawberries as compared to control. On Days 1-7, the percentage of gray mold incidence increased from 14.6% to 100% in control inoculated fruits, while there was was a maximum of 18.8% of gray mold incidence in 5 mg/clamshell- treated inoculated fruits observed on Day 6.

In uninoculated fruit, the percentage of gray mold incidence increased from 0% to 100% in control fruits, however, the percentage of gray mold incidence only increased to 54.1% in 5 mg/clamshell-treated uninoculated fruits.

Table 5. Ability of Compound A applied to clamshells by sublimation to control growth of Botrytis cinerea inoculated in strawberries.

Inoculated Fruit Gray Mold Incidence (%)

Treatment Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7

5 mg Compound A 0.0 0.0 0.0 0.0 0.0 16.7 18.8 16.7

Control 14.6 52.1 91.7 95.8 97.9 100.0 100.0 100.0

Gray Mold Severity (0-4)

Treatment Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7

5 mg Compound A 0.0 0.0 0.0 0.0 0.0 0.1 0.2 0.3

Control 0.1 0.4 0.9 2.0 3.0 4.0 4.0 4.0

Uninoculated Fruit Gray Mold Incidence (%)

Treatment Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7

5 mg Compound A 0.0 2.6 0.1 26.3 34.1 40.0 43.3 54.1

Control 0.0 23.0 0.6 95.2 100.0 100.0 100.0 100.0

Gray Mold Severity (0-4)

Treatment Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7

5 mg Compound A 0.0 0.4 1.0 1.0 1.0 1.3 1.3 1.6

Control 0.1 1.0 1.0 2.0 2.8 4.0 4.0 4.0 In addition, the 5 mg/clamshell concentration of active ingredient on sublimation- treated clamshells showed significant inhibition of gray mold severity in inoculated strawberries as compared to control (see Table 5). More specifically, on Days 1-7, the degree of gray mold severity increased from 0.1 to 4.0 in control inoculated fruits as compared to 0 to 0.3 in 5 mg/clamshell-treated inoculated fruits. For uninoculated fruits, the control fruits showed a gray mold severity level of 4.0 by Day 5, while the 5 mg/clamshell-treated uninoculated fruits only showed a gray mold severity level level of 1.6 on Day 7. Ultimately, this data demonstrate that treating clamshells with 5 mg/clamshell of benzoxaborole Compound A inhibited the gray mold infection of B. cinerea inoculated in strawberries.

Example 6: Benzoxaborole Compound Treatment to Different Locations of Fruit

Clamshells by Painting (In Vivo)

This in vivo assay was used to evaluate the ability of Compound A to volatilize from different locations of a clamshell (i.e., the base and/or the lid of the clamshell) in order to control or inhibit fruit infection by pathogenic microorganism, Botrytis cinerea. This experiment was conducted exactly as described in Example 1, with a few exceptions.

Compound A was administered to the clamshell by painting only. 5 mg of Compound A was painted onto the surface of the base of the clamshell or the lid of the clamshell (i.e., 5 mg/clamshell treatment rate). 2.5 mg of Compound A was painted onto the the base and the lid of the clamshell (for a total of 5 mg/clamshell treatment rate). The clamshell was then permitted to dry for 5 minutes. Strawberries were inoculated with Botrytis cinerea, placed within the clamshell for an initial time period of 5 days, and assessed for disease incidence and severity over a second time period of 7 days as described in Example 1.

The outcome of this in vivo experiment is summarized in Table 6. Results demonstrate good in vivo antimicrobial activity of Compound A against B. cinerea, with a reduction in disease incidence and severity with the 5 mg/clamshell concentrations applied by painting. In particular, both inoculated and uninoculated 5 mg/clamshell-treated clamshells showed inhibition of gray mold incidence and severity in strawberries as compared to control. On Days 1-7, the percentage of gray mold incidence increased from 0% to 100% in control inoculated fruits, while there was a maximum of 18.8% of gray mold incidence in 5 mg/clamshell base- treated inoculated fruits observed on Day 7. However, inoculated fruits in clamshells painted with 5 mg of Compound A on the lids only or the base and lids showed no incidence of gray mold even by Day 7.

In uninoculated fruit, the percentage of gray mold incidence increased from 0% to 100% in control fruits, however, the percentage of gray mold incidence only increased to 41.1%, 64.4%, and 52.2% in uninoculated fruits painted with 5 mg/clamshell on the base only, the lid only, and the base and lid, respectively.

In addition, the 5 mg/clamshell concentration of active ingredient on painted clamshells showed significant inhibition of gray mold severity in inoculated strawberries as compared to control (see Table 6). More specifically, on Days 1-7, the degree of gray mold severity increased from 0 to 3.6 in control inoculated fruits as compared to 0 to 0.3 in 5 mg/clamshell base-treated inoculated fruits. However, inoculated fruits in clamshells painted with 5 mg of Compound A on the lids only or the base and lids showed no increase in the severity of gray mold even by Day 7.

For uninoculated fruits, the control fruits showed a gray mold severity level of 4.0 by

Day 5, while the gray mold severity level level was 1.5, 2.5, and 1.5 on Day 7 in uninoculated fruits painted with 5 mg/clamshell on the base only, the lid only, and the base and lid, respectively. Ultimately, these data demonstrate that treating clamshells with/clamshell of benzoxaborole Compound A significantly inhibited the gray mold infection of B. cinerea irrespective of the location of the treatment application.

The preceding description enables others skilled in the art to utilize the technology in various embodiments and with various modifications as are suited to the particular use contemplated. In accordance with the provisions of the patent statutes, the principles and modes of operation of this disclosure have been explained and illustrated in exemplary embodiments. Accordingly, the present invention is not limited to the particular embodiments described and/or exemplified herein.

It is intended that the scope of disclosure of the present technology be defined by the following claims. However, it must be understood that this disclosure may be practiced otherwise than is specifically explained and illustrated without departing from its spirit or scope. It should be understood by those skilled in the art that various alternatives to the embodiments described herein may be employed in practicing the claims without departing from the spirit and scope as defined in the following claims.

The scope of this disclosure should be determined, not only with reference to the above description, but should instead be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled. It is anticipated and intended that future developments will occur in the arts discussed herein, and that the disclosed compositions and methods will be incorporated into such future examples. Table 6. Ability of Compound A painted on the base, lid, or base and lid of clamshells to control growth of Botryti cinerea inoculated on strawberries.

Inoculated Fruit Gray Mold Incidence (%)

Treatment Rate (mg)

Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Location

Base 5 0.0 0.0 12.5 12.5 0.0 0.0 18.8 18.8

Lid 5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Base and Lid 2.5/2.5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Control 0 0.0 25.0 56.3 68.8 93.8 93.8 93.8 100.0

Gray Mold Severity (0-4)

Treatment Rate (mg)

Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Location

Base 5 0.0 0.0 0.1 0.1 0.0 0.0 0.2 0.3

Lid 5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Base and Lid 2.5/2.5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Control 0 0.0 0.1 0.5 0.8 1.5 2.7 3.3 3.6

Uninoculated

Gray Mold Incidence (%)

Fruit

Treatment Rate (mg)

Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Location

Base 5 0.0 0.0 0.0 0.0 2.2 10.0 26.7 41.1

Lid 5 0.0 0.0 0.0 1.1 18.9 27.8 45.6 64.4

Base and Lid 2.5/2.5 0.0 0.0 0.0 1.1 7.8 23.3 33.3 52.2

Control 0 0.0 1.1 1.1 18.9 88.9 97.8 100.0 100.0

Gray Mold Severity (0-4)

Treatment Rate (mg)

Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Location

Base 5 0.0 0.0 0.0 0.0 0.3 0.8 1.2 1.5

Lid 5 0.0 0.0 0.0 0.2 1.2 2.0 2.0 2.5

Base and Lid 2.5/2.5 0.0 0.0 0.0 0.2 0.8 1.5 1.5 1.5

Control 0 0.0 0.2 0.2 1.3 3.7 4.0 4.0 4.0

Furthermore, all terms used in the claims are intended to be given their broadest reasonable constructions and their ordinary meanings as understood by those skilled in the art unless an explicit indication to the contrary is made herein. In particular, use of the singular articles such as "a," "the," "said," etc. should be read to recite one or more of the indicated elements unless a claim recites an explicit limitation to the contrary. It is intended that the following claims define the scope of the disclosure and that the technology within the scope of these claims and their equivalents be covered thereby. In sum, it should be understood that the disclosure is capable of modification and variation and is limited only by the following claims