NZ735129B2 - Concentrated natamycin suspension formulations - Google Patents

Abstract

The present invention generally relates to concentrated natamycin suspension formulations for inhibition of fungal growth. Specifically, the present invention relates to stable suspension concentrate formulations comprising from about 25 % to about 48 % w/w natamycin, from about 0.1 % to about 10 % w/w of an anionic surfactant being sodium lignosulfonate. and water. The formulations of the present invention contain natamycin as particles which are on average less than 11 microns in diameter and the formulations have a viscosity of less than 1400 centipoise at 21 degrees Celsius.

Description

CONCENTRATED NATAMYCIN SUSPENSION FORMULATIONS FIELD OF THE INVENTION The present invention relates to concentrated natamycin sion formulations for the inhibition of fungal growth.

BACKGROUND OF THE INVENTION Natamycin (syn. Pimaricin) is a e fungicide that is derived from the soil microorganisms, Streptomyces natalensis, Streptomyces lydicus, and Streptomyces chattanoogensis. It is commercially ed by fermenting and then lysing omyces natalensis. Natamycin is commonly used to deter the growth of fungus on edible solid foods, however, due to its poor solubility in water, it is often difficult to use.

One way the solubility issue has been overcome in the past is by the use of a wettable powder. A wettable powder formulation is a dry, finely ground formulation. In this type of formulation, the active ingredient is combined with a finely ground dry carrier, usually a mineral clay, and with other ingredients that enhance the ability of the powder to be suspended in water. Upon mixing the wettable powder with water, a suspension is formed, which is then applied by a spray technique.

One disadvantage of le powders is that the spray liquid must be continuously mixed to t settling of insoluble materials. Another disadvantage is that wettable powders and e powder formulations tend to produce dust upon handling, such as when pouring, transferring or measuring them. This dust may pose health hazards. r, powder formulations tend to wet poorly and also solubilize slowly upon addition to water.

Powder ations thus take longer to wet, disperse and solubilize in a tank-mix. Formation of lumps or partially solubilized spray solutions leads to uneven distribution of the cin in the tank-mix with the potential for reduced performance. Sometimes, foam in the spray tank caused by spray tank adjuvants can also affect wetting and solubility of wettable and soluble powders. Wettable powder formulations may also leave undesirable insoluble es both in the tank and on materials in need of treatment.

For many years those of skill in the art have attempted to p trated aqueous natamycin formulations. One challenge is that milling natamycin causes unacceptably high viscosity in most ations. If natamycin is not , however, suspension concentrates are not stable because the natamycin settles out of the ation. Yet another problem with natamycin formulations is that they are especially susceptible to bacterial growth.

U.S. Patent 5,552,151 suggests natamycin formulations with up to 40 % w/w natamycin.

This patent fails to provide any formulation examples with such high trations of natamycin. Further, the patent states that the most preferred range of natamycin is from 5 to 20 %. In addition, this patent fails to teach or suggest how to t the natamycin from settling out of a suspension concentrate formulation.

U.S. Patents 6,291,436 and 6,576,617 disclose solid natamycin formulations with particles less than 9 s in diameter. These patents, r, fail to teach or suggest how to overcome the viscosity issues associated with milled natamycin in liquid formulations.

DSM has successfully created suspension concentrate natamycin formulations with concentrations of up to 10 % w/w of natamycin (i.e., Zivion A (~4% SC), Zivion P (~4% SC), Delvo®Coat L02101 (~5% SC) and Delvocide L (~10% SC), Delvo is a registered ark of DSM). While these ations are successful, there is a need in the art for more concentrated formulations. These formulations will allow for reduced shipping and handling costs.

SUMMARY OF THE INVENTION ] In a first aspect, the present invention provides a stable suspension concentrate formulation for inhibition of fungal growth comprising: from about 25 % to about 48 % w/w natamycin; from about 0.5 % to about 2.5 % w/w sodium lignosulfonate; and water, wherein the natamycin is present as particles that on average are from 3 to 9 microns in diameter and the formulation has a viscosity of less than 1400 centipoise at 21 degrees Celsius. [0008b] In a second aspect, the present invention provides a stable suspension concentrate formulation for inhibition of fungal growth comprising: about 25% w/w natamycin; from about 0.5 % to about 2.5 % w/w sodium lignosulfonate; from about 0.1 % to about 10 % w/w of a nonionic surfactant selected from the group consisting of polyalkylene oxide block mers, polyoxyethylene sorbitan trioleates, polyoxyethylene sorbitol hexaoleates, and combinations thereof; and water, wherein the natamycin is present as particles that on average are from 3 to 9 microns in diameter and the formulation has a viscosity of less than 1400 centipoise at 21 degrees Celsius. [0008c] In a third aspect, the present invention provides use of the formulation of the first aspect or the second aspect in the manufacture of a ment for inhibiting the growth of a fungal pathogen in a subject in need thereof. [0008d] In a fourth aspect, the present ion provides a method of inhibiting fungal growth comprising: (a) diluting a suspension concentrate formulation which comprises from about 25 % to about 48 % w/w cin, from about 0.5% to about 2.5% w/w of sodium ulfonate, and water; and (b) apply ing the d suspension concentrate formulation to a commodity in need of protection from fungal growth, wherein the natamycin is present as particles that on average are from 3 to 9 microns in diameter and the formulation has a viscosity of less than 1400 centipoise at 21 degrees Celsius prior to dilution.

In another aspect, the present invention is directed to stable suspension concentrate formulations sing from about 0.1 % to about 10 % w/w of an anionic surfactant selected from the group ting of ectrolyte polymers, modified styrene acrylic polymers, dioctyl sodium sulfosuccinates, sodium salts of naphthalene sulfonates, and combinations thereof, and water, wherein the average particle size of the natamycin particles is less than 11 microns in diameter and the ation has a ity of less than 1400 oise at 21 degrees Celsius.

In a further aspect, the present invention is directed to methods for inhibiting the growth of human, animal, plant fungal pathogens or food spoilage fungi comprising applying or administering the formulation of the present invention to the human, animal, or plant in need thereof.

In another aspect, the present invention is directed methods for protecting a commodity comprising ng a suspension concentrate formulation which comprises from about 25 00 to about 48 00 w/w natamycin, from about 0| 00 to about 10 00 w/w of an c surfactant selected from the group ting of polyelectrolyte polymers, modified styrene acrylic polymers, dioctyl sodium sulfosuccinates, sodium salts of naphthalene sulfonates, and combinations thereof. and water, and applying the diluted suspension concentrate formulation to the commodity, wherein the natamycin is present as particles in the formulation the particles are on average less than I l microns in diameter and the formulation has a viscosity ofless than 1400 centipoise at 2| degrees Celsius prior to dilution.

LL‘D PTION 01" ll lL‘ l.\'\=’l;'.\"l'10\ Applicant unexpectedly discovered that using a specitic nonionic and anionic surfactant system, or using specific c surfactants, with cin particles of redttced size resulted in stable suspension concentrates with natamycin concentrations from about 25 0'0 to about 48 00 w/w. This ?nding was unexpected because numerous other surfactants, some with similar chemistries, failed to provide actory results (see for example, Example 13 below).

Applicant also found that the formulations of the present invention were not acceptable when they only included the tested nonionic surfactants because the viscosity was rable or too high (see lixample IO below).

As used herein, "suspension concentrate" refers to a formulation wherein insoluble particles are suspended in liquid or aqueous dilttents. A desirable characteristic of suspension concentrates is to have the insoluble particles evenly dispersed within the fortnulation. A suspension concentrate is not a solution.

] Applicant’s fortnulations allow for icant reductions in the costs of processing, packaging. storage, and transportation. The s formulations are also safer and tnuch easier to use than powdered fortnulations. In addition, all of the components of the formulations are approved by the US. linvironmental tion Agency for post-harvest use (see 40 Cl-‘R § 180.060) and are safe for use in or on food.

In one embodiment, the t invention is directed to stable suspension concentrate formulations comprising from about 25 0o to about 48 00 w/w natamyein, from about 0.1 00 to about 10 °o wiw of an anionic surfactant selected from the group consisting of polyelectrolyte polymers, modified styrene c polymers, l sodium sulfosuccinates, sodium salts of naphthalene sulfonates, and combinations thereof, and water, wherein the natamyein is present as particles that on average are less than I 1 microns in diameter and the formulation has a viscosity ofless than 1400 centipoise at 21 degrees Celsius.

As used herein, e" refers to a natamycin suspension concentrate 't'oz'mulation that does not form sediment or exhibit phase separation after being stored for a m of =18 hours at I degrees sétts.

The Viscosity of a ?uid is a measurement of the ?uids resistance to deformation by shear or tensile . l’luid Viscosity is frequently measured in centipoises (abbreviated as "cps") units with higher numbers correlating to thicker lluids. The viscosity of suspension trates must be under I400 cps to be desirable.

In a preferred embodiment, the formulations contain from about 30 00 to about 48 °o w/w natatnycin. In a more preferred embodiment, the formulations contain from about 35 °o to about 48 ° '0 w/w natamycin.

In yet another embodiment, the formulations contain from about 0.] °o to about 3.0 00 w/w of an anionic surfactant selected front the group consisting of poly-electrolyte polymers. modified styrene aetylic polymers, l sodium sullbsuccinates, sodium salts of naphthalene sulfonates, and ations thereof. In a preferred embodiment, the formulations contain from about 0.5 0o to about 2.5 0o of an anionic sulfactant selected from the group consisting of polyelectrolyte polymers, modified styrene acrylic polymers, dioctyl sodium sulfosuccinates, sodium salts of naphthalene ates, and combinations thereof. In a more preferred embodiment, the formulations contain from about 0.5 0o to about 1.5 0o of an anionic surfactant selected from the group consisting of polyelectrolyte polymers, modilied styrene acrylic polymers, dioctyl sodium sullosuccinates, sodium salts of naphthalene sulfonates, and combinations thereof.

In a preferred embodiment, the anionic surfactant is a polyelectrolyte polymer. In a more preferred embodiment, the polyelectrolyte polymer is a sodium lignosulfonate such as Ultrazine NA (available from Borregaard l'ech).

In another embodiment, the ations also contain from about 0.1 00 to about 00 w/w of a nonionic surfactant selected from the group consisting of kvlene oxide block copolymers, yethylene sorbitan trioleates, pol_\_='ox_\_'eth_vlene sorbitol hexaleates, and combinations thereof. In a preferred embodiment, the formulations contain from about 0. | 00 to about 3.0 00 of a nonionic surfactant selected from the group ting of kylene oxide block copolymers. polyoxyethylene sorbitan trioleates, polyoxyethylene sorbitol hexaleates, and combinations thereof. In a more preferred embodiment, the formulations n from about 0.5 00 to about 2.5 °o of a nonionic surfactant selected from the group consisting of ])ol_valk_\_-*lene oxide block copolymers, polymyethyletie sorbitan tiioleates, [)()I)-’()x_\-’etli_\»'lene sorbitol hexaleates, and combinations thereof. In a most preferred embodiment, the formulations contain from about 0.5 0o to about I5 00 of a nonionic surfactant selected from the group consisting of polyalk_\_-'Iene oxide block copol_\_-'mers, oxyethylene an trioleates. polyoxyethylene sorbitol hexaleatcs, and combinations thereof.

In a prefen'ed embodiment, the nonionic surfactant is a polyalkylene oxide block copolymer. In a more preferred embodiment, the polyalkylene oxide block copolymer is Atlasm (3-5000 (available from (‘roda (‘rop Care).

In another ment, the sion concentrate ations contain from about 46 ° 0 to about 75 ° 0 w/w water. In a more preferred embodiment, the formulations contain from about ()4 ° 0 to about ()7 ° 0 w/w watc r.

In yet another embodiment, the formulations contain natamycin as particles that are on average less than ll microns in diameter. In a preferred embodiment, the natamyein particles are on average less than or equal to 9 microns in diameter. In a more ‘ed embodiment. the natamycin particles are on e less than or equal to 5 microns in diameter.

In a most red embodiment, the natamyein particles are on average less than or equal to 3 microns in diameter.

In a further embodiment. the formulations contain an antifreeze agent. Examples of suitable antifreeze agents include ethylene glycol, s. |,2-propylene glycol, |,3-prop_v|ene glycol, I,2—butanedio|, l,3—butanediol, I,4—butanedio|, I,4—pentanediol, 3-methyl-l,5-pentanediol, 2,3- dimethyl—2,3-butanediol, trimethylol propane, mannitol, sorbitol, glycerol, pentaewthritol, |,4— c_vc|ohexanedimethanol, xylenol, and bisphenols such as bisphenol A.

In a preferred ernboditnent, the formulations contain frorn about I °o to about 10 00 wfw of antifreeze agent. In a tnore red embodiment, the formulations contain from about 3 00 to about 7 00 w/w of antifreeze agent. In a most red embodiment, the formulations contain from about 4 ° 0 to about 6 ° 0 w/w of antifreeze agent.

In a further embodiment, the formulations contain an antifoam agent, Examples of suitable antifoam agents include silicone based antifoam , vegetable oils, acetylenic glycols, and high molecular weight adducts of propylene oxide. One preferred antifoam agent is a silicone based oaming auent._.

In a preferred embodiment, the formulations contain from about 0.1 00 to about 5 0’0 W/JW of an antifoam agent. In a more preferred embodiment, the formulations contain from about 0.5 °o to about 2 °o w/w of an antifoam agent. In a most preferred embodiment, the formulations contain from about 0.8 0 o to about I .2 ° 0 w/w ofan antifoam agent.

] In a further embodiment, the formulations contain an antibacterial preservative. es of suitable antibacterial preservatives include benzoates and K-sorbate.

In a preferred embodiment, the formulations contain frotn about 0 01 ° 0 to about 3 °o w/w of an antibacterial vative. In a more preferred embodiment, the ations contain from about ().I °o to about 2 °o w/w ofan antibacterial preservative, In a most preferred embodiment. the ations contain from about 0.3 on to about I °o w/w of an antibacterial preservative.

] In a preferred embodiment, the formulations n: about 25% w’w natarnycin; from about 0.1 "o to about lO 00 w/w of an anionic surfactant selected from the group consisting of polyelcctrolytc polymers, modified styrene acrylic polymers, dioctyl sodium sulfosuccinatcs, sodium salts of naphthalene ates. and combinations th ereof; from about ().l "o to about 10 0o \vfw of a nonionic surfactant ed from the group consisting of polyalkylene oxide block copolvmers, polyoxyethylene sorbitan trioleates, polyoxyethylene sorbitol hexaleates, and combinations thereof; and water, wherein the natamycin is present as particles that on average are less than I | microns in diameter and the formulation has a viscosity of less than 1400 centipoise at 2] degrees Celsius.

In a more preferred embodiment, the formulations contain: about 25° 0 w/w natamycin; about 1.0% w/w sodium lignosulfonate; and about 0.50 0 w/w ofa polyalkvlene oxide block mer, wherein the nalamycin is present as particles that on average are less than I l microns in er and the formulation has a viscosity of less than I400 oise at 2| degrees Celsius.

In another embodiment, the present invention is directed to methods for inhibiting the growth of human, animal, plant fungal pathogens or food ge fungi comprising applying or administering the formulation ofclaim l to the human. animal, or plant in need thereof.

In a preferred embodiment, the formulations of the present invention are used to inhibit fungal pathogens by administration of the ations to a human. Suitable uses include treatment of fungal infections in the eyes, mouth or on the skin (fungal tis). Preferably, the formulations are administered directly to the area in need of fungal growth tion. The formulations are suitable for control of du. Aspcrgi/lns, ('cp/u[Ins/2017111", I’lls'ul'ium, and Penicillin/n growing on a human.

In another preferred embodiment, the formulations of the present invention are used to t fungal pathogens by stration of the formulations to an animal. Suitable uses include treatment of fungal infections in the eyes and the surrounding tissues and ringworm infections. Suitable animals include domesticated animals, such as dogs, cats, cattle and horses.

Preferably, the formulations are administered ly to the area in need of fungal growth tion. The lbrmulations are suitable for control of ( ‘(mc/I'cla, Aspergi/lus. ( 'ep/m/os/)ormm, lv'IIs‘ur/‘mn, and Penicillin/n growing on an animal.

In another preferred embodiment, the formulations of the present invention are used to inhibit fungal pathogens by administration of the formulations to a plant. Suitable plants e commercially cultivated crops.

In a further embodiment, the tormulations of the present invention are used to inhibit mold and yeast growth in mushroom growing medium.

In a preferred embodiment, the suspension concentrate ations that are used for inhibiting the growth of human, animal, plant fungal pathogens or food spoilage fungi further comprise from about 0| 00 to about IO °o w/w of a nonionic surfactant selected frotn the group consisting of polyalkylene oxide block eopolymers, polyoxyethylene sorbitan trioleates, polyoxycthylcne ol hexalcates, and combinations thereof.

In another ment, the present invention is directed to methods for inhibiting fungal growth comprising diluting a sion concentrate formulation which comprises front about 25 00 to about 48 °o w/‘w natamycin, from about 0| 00 to about 10 00 w/w of an anionic surfactant ed from the group consisting of ectrolyte polymers, modified styrene acrylic polymers, dioctyl sodium sulfosuccinates, sodium salts of naphthalene sulfonatcs, and combinations thereof, and water, and applying the diluted suspension concentrate formulation to a commodity in need of protection frotn fungal growth, wherein the natamycin is present in the formulation as particles and the les are on average less than 1 1 microns in diameter and the formulation has a viscosity ofless than 1400 cenlipoise at 21 degrees (‘elsius prior to dilution.

In a red embodiment, the suspension concentration lbrtnulalions that are used for inhibiting fungal growth r comprise from about ().| "o to about 10 00 w/w of a ic surfactant selected from the group consisting of polyalkylene oxide block copolymers. ox_veth_\"lene sorbitan ates, polyox_\-’eth_\;‘|ene sorbitol hexaleates, and combinations thereof.

In an embodiment, the formulations of the present invention are applied to the ity by dipping, drenching or spraying.

In another embodiment, for formulations are diluted to from about 0.01 0o to about 5 °o w/w natamycin before being applied to the commodity. In a preferred embodiment, the formulations are diluted to from about 0.5 "a to about 2 0’0 w/w natamycin before being applied to the commodity, WOO-"33] in an embodiment. the commodities inciude cheeses, stsiccs fruits, vcgetz bias, nuts, cereal grains, animal feed. spices. ges or other products intended for consumption. i The formulations are suitable for use on a y of {"LCSCS The iormulntions of the present invention may be added to r disparsirms used to eitcase the cheese (such as a rind) or the cheese may be dipped or sprayed with the formuiations ofthc titrcscnt invention. The forii'iulations may aiso be added to shredded cheeses le cheeses include Gouda. cdam, cheddar, tilsitcr: caciotta. fontina, taliegio. montasio, axingc provolone. pcccn‘ino, romano: blue checscs. and indian cheeses. littttiatSE 'l‘hc- formulations are suitable for use on a variety of sausages. 'i'iic formuiations of the present in'venticm may be used to dip or spray the outside of the es. iixltei'iiatively, the sausage: casings may he treated bct‘i‘irc they are titled. The lormulations may aiso be air-plied to fermented meat produczs. Suitable es include Dutci raw sausages, German raw sausz-igcs. and ltalian es The formulations are suitable for use on many types of fruits and vegetables. The formulations of the present inVention may be dispersed onto the fruits while they are growing, just prior to harvest. or after harvest. In a red embodiment, the formulatimts of the present invention are dispersed onto the fruits and vegetables after they are harvested. In an alternative embodiment, the formulations of the present invention are dispersed onto the fruits and vegetables while they are still growing on the plants. As used herein, ted" or "post- haryest" means that the fruits or vegetables have been d from the plant they grew on and are no longer growing. As used herein, "dispersion" refers to evenly distributing the formulation over the commodity surface.

As used herein cting" refers to reducing the likelihood of fungal pathogens induced rot.

In a preferred embodiment. the suspension concentrate formulation that is applied to the fruits or vegetables also includes from about 0.1 °o to about lO 00 w/w of a nonionic tant selected from the group consisting of polyalkylene oxide block copolymcrs, polyoxyethylene sorbitan trioleates, polyoxyethylcne sorbitol hexaleates, and ations thereof.

Many types of agriculturally or pharmaceutically acceptable diluents may be used to dilute the lormulations of the present ion. l-‘or example, water, glycerol, hexylene glycol, dipropylene glycol, and poly-ethylene glycols are all acceptable diluents.

As used herein "fruit" refers to the ?eshy tissue associated with a seed of an edible plant. Examples of fruits include citrus fruits, berries, pome fruits (such as apples), stone fruits, melons and bananas ] Examples of citrus fruits include oranges, grapet‘ruits, clernentines, ins, limes, pornelo, kumquats, and hybrids thereof.

Examples of berries include grapes. aronia berry, bayberry, bearberry, bitberry, blackberry, blueberry, lowbush blueberry, highbush blueberry, buffalo currant, buffaloberry, che, Chilean guaya, erry, cloudberry, cranberry, highbush cranberry, black currant, red currant, elderberry, L-‘uropean ry, gooseberry, grape, edible honeysuckle, huckleberry, jostaberry, Juneberry. lingonberry, maypop, mountain pepper berries, mulberry, es, native t. partridgeberry, plralsa, pincherry, black raspberry, red raspberry, , salal, schisandra berry, sea buckthorn, serviceberry, strawberry, wild raspberry, and ars, varieties and s thereof.

] Examples of pome fruits include apple, azarole, ple, loquat. mayhaw, medlar, pear, Asian pear, quince, Chinese quince, Japanese quince, tejocote, and cultiyars, varieties and hybrids thereof. lixarnples of stone fruits include apricot, sweet cherry, tart cherry, nectarine, peach, plum, Clricksaw plum, Damson plum, Japanese plum, t. fresh prune, and cultivars, varieties and hybrids thereof.

Examples of melons include Citron , muskmelons, watermelon, cantaloupe, casaba, crenshaw melon, golden perslraw melon, ew melon, honey balls, mango melon, Persian melon, pineapple melon. Santa (‘laus melon. snake melon and cultivars, varieties and hybrids thereof.

] In one embodiment, the d formulation is applied to citrus fruits to protect them from sour rot. "Sour rot" refers to a fungal infection caused by ( iconic/mm cilri—um'umii.

In another embodiment, the diluted formulation is applied to citrus fruits to protect them from "green mold" or "blue mold." 'l'hese molds refer to a fungal infection caused by Penicillin/n m) or Panic/Ilium [It'll/CHI".

As used herein "vegetable" refers to root and tuber vegetables, bulb vegetables, leafy assica vegetables, leafy brassica vegetables, succulent or dried legumes, fruiting vegetables, and cucurbit vegetables.

In a preferred embodiment, the diluted formulation is applied by dipping, drenching, or spraying the commodities with and without coating waxes. in another embodiment, the formulations are diiuterj to a concentration of from about 0.0l 0'0 to about 5 00 w/w cin before being applied to the ities. In a preferred embodiment, the formulations are diluted to a concentration of from about 0.5 00 to about 2 0’0 w/w natamycin before being applied to the commodities.

] Examples of beverages include juices, beer, wine, soft drinks, iced tea and fruit s. Suitable juices include fruit juices and blend thereof, including lemonade and orange juices. {00062: The formulations of the present iim couid also include the fungicides lucensomycin. nysiatin, anaphetericin—B or combinations thereof. t00063] the formulations of the n invention speci?cally exclude thickening agents and anti-eating als. $00054] 'fhe formulations ofthe present invention speeiiicaliy exclude lactose powder and sodium chloride. ]€3{3065] The formulations of the present irn-cntion ically exclude polyoxzyethylenc alcohol nonionic surfactants, poiyerqfetliylene lanryl ether nonionic surfactants. and p<3lyxnyetiaylene cct‘yl ether rionicmic surfactants $60066] The formulations of the present inantion may he applied so that the natamycin concentration is less. than l ppm or greater than 5.- ppm. |00067| As used , all numerical values relating to amounts‘ weight percentages and the like, are defined as "about" or "approximately" each particular value. plus or minus IO 0o.

For example. the phrase "at least 5.0 00 by weight" is to be understood as "at least 4.5 °o to 5.5 00 by weight." 'l'herefore, amounts within 10 0'0 of the d values are encompassed by the scope of the claims. 00068 The disclosed embodiments are sim )lv exem )larv embodiments of the ivel - - concepts disclosed herein and should not be considered as limiting, unless so stated.

The following examples are intended to illustrate the present invention and to teach one of ordinary skill in the art how to make and use the invention. They are not intended to be limiting in any way.

EXAMPLES .Applicant used natamycin in the form of Technical Grade Active Ingredient ("TGAI") when preparing the formulations of the present invention. The percent cin in the technical grade typically ranges between 80 "o and 00 00 w/w. Variations in the activity of natamycin in the 'l'GAl should be accounted for by decreasing or increasing the amount of diluent in producing the natamycin formulation of the desired concentration This is rd practice within the guidelines of US Environmental Protection Agency per 40 CPR. § 158,175(b)(2). |0007l| Applicant prepared the following cin suspension concentrate ations as follows unless otherwise noted. .\'atamycin was sed until it was on average below 1 1 microns in diameter. The other formulation ents were added and the formulation was stirred until the natamyein particles fully dispersed. "Qs." refers to a sufficient quantity of water to reach the 1)1'0)eI'00 w/w of the formulation.

Example I Concentrated 25 0’0 Natamvcin Sus ension Formulations Table 1 Com )ouent (all in WW1) "mm \atamvcin technical wwdc-r Pol -‘r'alk ilene oxide. block-‘ . . . 0.5-2.5 0.5-2.»- co )olvmer nonionic surfactant l’olyoxycthylene sorbitan 0 5 7 S trioleate nonionic tant " H" l’olyoxyctltylene sorbitan hcxaoleatc nonionic surfactant Sodium lignosulfonate (polyelectrolyte pol y mer anionic surfactant) d styrene acrylic )olvmer anionic surfactant l sodium sulfosuccinates c surfactant Sodium salts of naphthalene sulfonatc anionic surfactant .\ntibacterial .rrcscrxatixc ----—- ——--— Example 2 Additional Concentrated 25 0o \atamvcin Sus cnsion Formulations Table 2 Com )onent (all in °o wi/wi) \atamvcin technical owdcr Polt-ialkylcnc oxide block copolymcr nonionic __ surfactant '- Sodium Ii gnosulfonatc lcctrolytc polymer anionic surfactant) ' ' Antifreeze agent Antibacterial )i'cscr'vati\-'c 0.3-] Antifoam agent 1 | Water ( .s‘ ( is. |00°o 100% Example 3 (Tonccntratcd 35 ° 0 Natamycin Suspension Formulation Table 3 \atarmcin technical — Antifreeze a cut l’olyalkylene oxide block (:0 olymer nonionic surfactant Sodium lignosull'onate c ——\ntiba'ctcrial )rcscnativc —100 °o Example 4 Concentrated 40 oh Natamvcin sion Formulation Table 4 % fit/wt l’olyalkyletieoxide block co olymer nonionic surfactant Sodium lignosull‘onate anionic surlactanl AllllbaClCl1211 )ICSCI‘V‘allVC Antiloam agent Water —100 "0 Example 5 Concentrated 45 ° 0 Natam3’Cll] Sus n Formulationl Table 5 \atamxein technical — Antifreeze a ’cnt — l’olyalkylene oxide block co olymer ic surfactant Sodium lignosull'onate anionic surfactant ——\ntiba'etei'ial )iesenative —100 °o Concentrated 46 oh Natamvcin Suspension Formulation Table 6 % "t/wl l’olyalkyleiieoxide block co olymer nonionic surfactant Sodium lignosull‘onale anionic surlactanl AllllbaClCl1211 )ICSCI‘V‘allVC Amiloam agent Water —100 "0 Example 7 (‘onccntratcd 47.5 °o Natamycin Suspension Formulation Table 7 \atzinwein technical l’olyalkylene oxide block co olymer nonionic tant Sodium lignosull'onale anionic surfactant ——\nliba'clei'ial )iesenativc —100 °o Comparative Example 8 trated 50 ob l\'atamvcin Suspension Formulation Table 8 % til/wt .\atam\ein technical Anm‘rccm went — Polyalkylone oxide block co olymer nonionic surfactant Sodium ull‘onate anionic surfactant Antibacterial vative Antil‘oam agent 1 Water ( _s. —100 "0 Example 9 Viscositv Studies ] Applicant tested the viscosity ol‘thc formulations of Examples 2A, ZB, 5, 6, 7 and Comparative Example 8. This study was conducted using standard procedures known and accepted by those of skill in the art. The results of this study can be seen below in Table 9.

Table 9 Formulation % w/w \alzlmycin Viscosity (cps) Example 2A 2 1080 ——Example 2B Example 6 040 Example 8 47.5 1375 |00073| As can be seen in Table 0 above, the compositions of Examples 2A, BB, 5, 6, and 7 haye viscosities that render the compositions suitable for use as a suspension concentrate for administration to plants, animals, or humans sities were less than I400 cps) and for other uses which has similar viscosity restrictions. In contrast, the 50 00 natamycin formulation of Comparative e 8 was unacceptably thick (2225. cps). Accordingly. Applicant detennined that the ations at concentrations of up to 48 ° 0 natamycin are suitable. e If) Additional \-"iscositv Studies [00074| Applicant tested the viscosity of 25 "o natamycin suspension concentrate ations with 2 °o of ent nonionic surfactants. This study was conducted using standard procedures known and accepted by those of skill in the art. The results ofthis study can be seen below in Table IO.

Table 10 Surfactant (2 "/0 w/w) Viscosity (cps) Control, water added instead of surfactant Control, anionic surfactant, sodium | i gn osul fonate Polyoxyethylene (20) sorbitan monolaurate, nonionic surfactant Acrylic copolymer, nonionic tant Polyoxyethylene (20) oleyl ether, nonionic surfactant Polyalkylene oxide block copolymer, nonionic surfactant Polyoxyethylenc (20) an trioleatc. nonionic surfactant Polyoxyethylene (40) sorbilol hexaoleate, nonionic surfactant As seen in 'l‘able [0 above, all of the six nonionic surfactants tested failed to provide the desirable viscosity (-~l400 cps or lower) when used alone in a ’25 00 suspension concentrate natamycin formulations. In contrast, the anionic tant, sodium ligm)sulfonate, used alone provided the desirable viscosity in a 25 00 suspension concentrate natamycin formulation.

Example ll Particle Size Stability" Study Applicant tested how the particle size of natamycin impacted the stability of suspension concentrate formulations. This study was conducted using standard procedures known and accepted by those of skill in the art. A 25 00 natamycin formulation with natamycin particles with e (I)(4,3)) diameters of | 1 microns v'as compared with a 25 0o natamycin formulation with natamycin les with average diameters of 3 microns. The results of this study can be seen below in Table l I .

Table 11 Average Initial 30 mins 24 hours 48 hours |)(4,3) Natamycin Particle Diameter (microns) Homogenous Some 1 (3 ° 0 48 ° 0 VA sedimentation sedimentation ntation I Iomogenous I Iomogenous; I nous; I Iomogenous; I nous; no no no no sedimentation sedimentation sedimentation sedimentation or separation or separation or tion or separation As can be seen in Table l 1 above. the ation with the smaller natamycin particles was homogcnous and did not exhibit sedimentation or separation even after 48 hours.

In contrast. a similar formulation with larger natamycin panicles formed sediment after only 30 minutes. By 24 hours, there was ive sedimentation that required IO inversions to re— suspend.

This study shows that particle size is an important aspect of ant‘s suspension concentrate formulations Example 12 Another Particle Size Stability Studv Applicant tested how particle sizes of 9 microns impacts the stability of the suspension concentrate formulations. The study used standard procedures known and accepted by those ot‘ skill in the art A 25 00 natamvcin formulation with natamycin particles with average (l)(4,3)) diameters of II microns was compared with a 25 l’o natamycin formulation with ein particles with average diameters 01") microns. The results 01' this study can be seen in Table 12 below. 'l'able 12 e Initial 30 mins 4 hours 24 hours 48 hours Natamycin Particle Diameter (microns) llomogenous Some 16 ° 0 48 ° 0 N.-"A sedimentation sedimentation sedimentation IIomogenous IIomogenous IIomogcnous 1.4 "o 2.80 o Sedimentation ntation As can be seen in Table 12 above, the formulation with the smaller natamycin particles was homogenous and did not exhibit ntation or separation after 4 hours. In contrast, a similar formulation with larger natamycin les formed sediment after only 30 minutes. By 24 hours, there was excessive sedimentation that required 10 inversions to re- suspend, | This study shows that particle size is an ant aspect of Applicant’s suspension concentrate formulations.

Example 13 Testing of :'\lternatiye .\'onionic and Anionic Surfactants While developing the formulations of the present invention, Applicant also tested numerous other surfactant combinations that failed to show synergy. A summaty of these results is below in Table I3.

Table I3 Nonionic Surfactants I; :- :75 U, 3 2 <1) 3 , T 2 E :2 L" v: a a a, .= z o 2 é a, 2 o o o 3 C E <1) :5 o g '3‘. 3‘. '3 8. 2 2' :1 6 3‘. a; E :E E o <33 52 E ': E 2 u *— — . E’ S’ E) . u, 3". 5h 9} 9 Anionic ;‘ 3 Q Q .2 :4 o E" § -§ E: 9 s 9 9 E 96 9 .: 9 .: 8 .2-. 2 ‘ a w. ‘zs .5 k. _Q 7-».

Surfactants _Q 3 3 5 U E 2 _o ,5 3 5 o. E o. o. < A. _c a. :1) a. :1) Polyelectrolyte i\'o l\‘o No No Synergy Synergy Synergy polymers synergy synergy synergy y Modi? ed styrene No l\' o z\' o Xo Synergy Synergy Synergy c polymers synergy y synergy y Doctyl sodium No No ;\'0 .\'o Synergy Synergy sulfosuccinates synergy synergy y synergy Sodium salts of :\'o l\'o :\'o \o Synergy Synergy Synergy naphthalene synergy synergy synergy synergy sulfonates As illustrated in Table 13, Applicant was unable to predict which surfactants would exhibit synergy and allow for large amounts (up to 48 00 w/w) of natamycin to be suspended while providing the desirable viscosity of the formulations. Applicant tried polyoxvethvlene alcohol nonionic tants, polvoxyethylene lauryl ether ic surfactants, polvoxvethvlenc cetyl ether nonionic surfactants. and acrylic copolymer nonionic surfactants without success. In contrast, the claimed surfactants all provided suitable formulations that were stable and had low viscosity. ol y Mold and Blue .V'lold ol~ Apple [00084| Formulation 2B from Table 2. above. was tested for its ability to control gray mold and blue mold of apple.

Method [MUCH/Ill)! preparaI/on One isolate each of Bally/11v c’illt’l't’t'l (A810) and llin/n mpcmsmn (A003) was recovered from stock culture collections saved in silica-gel at 4°C. After 3 days of growth on potato dextrose agar (PDA). I}. cine/'ca was sporulatcd under a lZ-?uorcsccnt light for 2 weeks. A conidial suspension was made by adding 20 ml of sterile water and gently removing the conidia with a sterile c loop. A spore suspension was filtered h two layers of cheesecloth. The final tration was ed to l / IOi conidia/ml with a hemacytometer.

For I’. expuns'um, dry conidia were transferred into 5 ml of sterile water containing 0.01% Tween ® (Tween is a registered trademark of Croda Americas 1.1.0 by a plastic loop after wetting in the Tween 20® water. After vortexing vigorously, spores were diluted in 0.01% Tween 2069 water and adjusted to I /- IO'l conidia/ml. l'i'm'l inoculation Organic 'Red Delicious’ apples that were washed in hypochlorite and packed in a commercial apple packinghouse were purchased. The apples were incubated at ambient temperature at least 12 hours prior to the experiment. The apples were then wound-inoculated by adding IO pl ofthe inoculum suspensions prepared as above into a wound created with a finished nail-head (3 x 4 mm). The apples were incubated at room temperature for 4 hours before fungicide treatments.

[Q Is) [real/Hem The apples were placed in a polyethylene mesh bag and dipped in fungicide solutions for 30 seconds. Fungicides tested were Formulation 2B of the instant invention, ZivionCR) .Vl ( 10.3400 natamycin, Zivion is a registered trademark of and available from DSM ll’ Assets B.\". of the Netherlands) and Shield—BriteCR) FDL 2308C (20.4% lludioxonil, Shield Brite is a registered trademark of Pace International, LLC; Shield-Brite® FDL 2308(‘ is available from Pace International, l.l.C). After treatments, the apples were placed on fiber apple trays in a cardboard box, and stored at 4"(‘ in air. Apples dipped in water were used as a control. Twenty apples per replicate and 4 replicates per treatment were used. At least l-Kg apples per treatment were treated and used for fruit e analysis. liquid ons were sampled and analyzed for natantycin trations. 1. Mia (Illa/J "xix The tions were conduct after 6 weeks of cold storage, The percent ofdecay incidences in the dip ents were arcsirte-transformed and ed with SAS l’ROC‘ GLM (version c),|; SAS Institute) to compare the treatments. Means were separated by Fisher‘s protected least significant difference at l’ — 0.05.

Results This study was conducted to determine the effectiveness of natamycin formulations of the present invention in controlling (ray moldha and blue mold of apple by aqueous dip ation.

In general, Formulation 2B of the instant invention showed better performance than Zivioni?) \l in controlling both gray mold and blue mold of apples when d by an aqueous dip. See 'l‘ables l4 & l5. respectively. Formulation 2B was equally effective as fludioxonil 180 ppm against both pathogens when applied at 750 ppm or 1,000 ppm. All 3 rates of Formulation BB were equally effective to each other in controlling gray mold (Table I4), whereas SOO-ppm Formulation 2B was less ive than 750 ppm or 1,000 ppm in controlling blue mold (Table 15).

Regardless of the rates, apples treated with '/.ivion® M showed higher decay incidences ofgray mold and blue mold than those treated with Formulation 2B. Surprisingly. the l\) Lu difference in decay rate cannot be explained solely by the 2.5 times difference in the amount of cin between Formulation QB and Zivion® NI. First. Formulation 28 showed more than a 2.5 times reduction in mold oyer Zivion® M. Speci?cally, at an application rate of 1.000 ppm Formulation 213 resulted in 3.5 times less gray mold than application ofthe satne rate oniyion® .\I. See [able [4 At an application rate 01 750 ppm lomtulation 2B resulted in 43 times less gray mold than Ziyion® .\»1. At an application rate of 750 ppm Formulation 213 ed in 3.68 times less blue mold than Ziyion® M. [00092| Secondly, the liquid concentrations and fruit residues of natamycin were very similar between Formulation EB and Ziyion® .\-'l in both trials. See Tables 14 & l5. Thus, the natamycin formulations of the present ion show unexpected results over commercially available ttatamycitt lormulalions.

] Further research will be needed to investigate why the natamycin formulations performed dill‘erently on the control y mold and blue mold in apples.

Table 14 .Vleasured natamycin ( ) mt) Treatment —-_mz_ Formulation 213 Zivion® M Shield-Brite® FD], 230sc * Values with a common letter are not cantly different according to the analysis of variance and least significant difference at l’ — 0.05. 'l'able IS Measured natamycin 'l‘qr a rate‘ g ent In Blue mold (%) ("W") —_——— ation 2R Shield-Bruce) FDL 23080 Values with a common letter are not signi? cantly different according to the analysis of variance and least significant difference at l’ — 005.

Example IS (‘ontrol ol‘ Postharvest Diseases in Citrus l-‘ormulation QB from Table 2, above, was tested for its ability to l green mold of lemons and oranges and sour rot in lemons.

Methods I'l'uil Preparation Two bins each ol‘light green 'Eurcka" lemons and 'Washington navel" oranges were harvested December 15, 2014 at the sity ol‘(‘alit‘ornia, Lindcove Research and Extension r (UC—LREC). An additional bin ol‘vellow. more mature ‘Eureka" lemons was provided by Pace ational, LLC that was harvested from District I region of California. All fruit was pressure washed at 100 psi for 20 seconds in 200 ppm chlorine. then divided into totes of 150 fruit each. The totes were placed at 20"(‘ for 36 hours before ation from 6 to 9 PM. on December l7, 20l4. [noon/um preparation One isolate each ol‘l’cn/cri/lium dig/lam"; (M557; fungicide sensitive) and (ico/r/c/mm cil/‘i-um'anli/ (A005) was used for this study. Fungal cultures were reactivated from silica-gel stocks and grown on PDA The plate was ?ooded with 0.01% 'l'ween® '20 solution for l’. digi/ulmn and sterile water for (j. cill'i-uumnlii, and conidia were scraped off with a sterile loop. The ?nal concentrations were adjusted to 5 " I05 and 5 * IO- sporest’ml for I). dig/"lulu": and ( i. uumnlii, respectively. Both ons were kept in an ice chest until they were used. 1'7?!" ation ] Lemons and oranges with no postharvest fungicide ents were used. l"or 1’. ") and (i. ciII'I—(mrcm/ii inoculation, a sterile steel rod ( l x 2 mm) was dipped in the inoculum suspension and fruit was wound-inoculated by making a single puncture on equatorial surface I2 to 16 hours before treatments were applied. The pallets oflemons inoculated with (1. ciIri-um'cmlii were covered with a plastic bag to maintain high humidity. After treatment. fruits were placed on fruit cavity trays in cartons and placed in the large temperature-eontrolled room at LIC-IRFC at 50"F.

Fungicides tested were Formulation 2B ol‘the instant invention, Shield-Brite<® I’DL 2308C, \"lentor(R) (45% propiconazole; Mentor is a registered ark and available from Syngenta l’articipations AG). Shield-Brite® TH] 5000 (42.300 thiabendazole; available from l’ace International, l.l.C‘), Shield-Brite® l’enbotec® 4005C (37. I400 ntanil; l’enbotec is a registered trademark ol‘Johnson & Johnson Corporation; -Britcdi; Penbotec® 4008(‘ is available through Pace International, LL(‘). PacRite® lor 75 WSG ( 10000 imazalil; PacRite® is a registered trademark e International, LLC; PacRite® Fungal‘lor 75 W80 is available through Pace International, LL(‘). and Graduate A | (R) (l'ludioxonil in combination with azoxvstrobin; Graduate A | is a registered trademark of and available through Syngenta Patticipations AG).

Application A lei/20d [00099| Fungicides were applied by recirculating ?ooder with two s and a residence time ofabout 7 seconds with a system volume of 75 gallons and a flow rate of 50 gal/minute with or without an addition of coating wax, except that thiabendazole was applied by spraying with a pack wax in wax cabinet with dryer at l20°l-‘ ("pack wax").

Y ‘I‘emmcm 1. Green mold on lemons [000I00| Fruit were inoculated l2- I 6 hours before treatment at 68°F with l’. digital/um.

Treatments were applied to 3 sets of45 lemons from 0 AM. to 2 PM. on December IS. 2014 followed by storage at 5( "F until J anuary 5. 2015 when they were examined for decay development. 2. Green mold on oranges Fruit were ated 12-16 hours before treatment at 68°F with 1’. dig/"(alum ’l‘reatments applied to 3 sets of 45 oranges from 9 A..\l. to 2 PM. on December 18. 2014 ed by storage at 50°F until January 5, 20 I 5, then 3 days at 72°F, then 4 days at 50°F that ended on January 12, 2015 when they were examined. 3. Sour rot on lemons Fruit were ated 12-16 hours before treatment at 68°F with (j. cilri-uHrun/ii.

Treatments applied to 3 sets 01°45 lemons from 9 AM. to 2 PM. on December 18. 2014 followed by storage at 50°F until y 5, 2015, then 3 days at 72°F, then 4 days at 50°F that ended on y 12, 2015 when they were examined. 1 Mia unabxvix The t ofdecay incidences were arcsirte-transformed and analyzed with SAS PROC (3134 (version 0.1; SAS Institute) to compare the treatments. Means were separated by Fisher‘s protected least significant difference at l’ — 0,05. The mean separation via Fisher’s Protected LSD)": in Table 16 below was 3.4 for green mold on lemons. 13.6 for green mold on oranges and 5.8 for sour rot on lemons.

Results This study was conducted to determine the effectiveness of natamycin formulations of the present invention in controlling green mold of lemons and oranges and sour rot of lenrorrs by aqueous dip or llooder application.

In general. application of Formulation 213 of the present invention ed in significantly less green mold and sour rot than control. See Table 16. Further, ation of Formulation 2B resulted in statistically less green mold and sour rot than commercial fungicides.

Specifically, application of Formulation 2B via a tlooder at 500 ppm and via a flooder at 1.000 ppm with storage was resulted in statistically less green mold on lemons than 2.000 ppm Shield- Brite® 'I'BZ 5001) applied via a spray with pack wax. See Table 16. ation of Formulation 21% via a ?ooder at 500 ppm and 1.000 ppm and via dip at 500 ppm resulted in statistically less green mold on oranges than 300ppm Shield—Brite® l-‘l)l. 2308C applied via a ?ooder. See Table . Finally, ation of Formulation 21-3 via a ?oodcr at 500 ppm and 1,000 ppm resulted in statistically less green mold on oranges than 3,500 ppm SlIield-Brite® 'l‘BZ 500D applied via a spray with pack wax. See Table 16. In conclusion, 1-‘ormulation 2B of the present invention is capable of lling green mold and sour I‘Ol 0n lemons and oranges as well as, and in some examples better than, commercially available fungicides.

Table 16 ’l‘arget"rate Application GI'H‘" "WM ("/0) ml ("/0) ent \Iethod . m— '—_-EI96.3 37.8 Formulation ZR 500 Floodcr l’onnulation 213 500 Dip 30 sec 20. l 4.4 Formulation 28 1000 Flooder 22.4 14.7 Storage l’ormulation 2B 1000 |"|ooder wax. no Pack wax, Fonnulation 2B Floodcr no fungicide Formulation ZB Dip 30 sec Shield-Britc® FDl. l-‘loodcr -3OSC Shield-Brite?s‘: FDI. lilooder 230SC PacRite® Fungal‘lor l-‘looder 75 SWG Shield-Britc® Floodcr Penbotcc® 4008C Sliicld-Brite® TBZ , Pack wax, 5001) ‘ ‘ ' fungicide Formulation '28 + Shield-Brite® l"1)1. 500 | 180 |"|ooder 2308C 1-‘ormulation 213 | -Brite® HM, 1000 ' 300 lilootler 2308C Formulation 213 | PacRite® Fungaf‘lor 1000 - 500 Floodcr 75 SWG formulation 213 | Shield-Blilc® 1000 ‘ 500 Floodcr Penbotec® 4008C ation EB + Pack wax Shield-BIiIeCR IBZ 1000I 3500 213 ?oodel l 5 fungicide emanate .x—o 300+ «,00 Me, —n— x-Iemmao 540 Home] ——— 1 540 we ———

Claims (16)

1. A stable suspension concentrate formulation for inhibition of fungal growth comprising: from about 25 % to about 48 % w/w natamycin; from about 0.5 % to about 2.5 % w/w sodium lignosulfonate; and water, wherein the natamycin is present as particles that on average are from 3 to 9 microns in diameter and the formulation has a viscosity of less than 1400 centipoise at 21 degrees Celsius.

2. The formulation of claim 1 further comprising from about 0.1 % to about 10 % w/w of a ic surfactant ed from the group consisting of polyalkylene oxide block copolymers, polyoxyethylene sorbitan trioleates, polyoxyethylene sorbitol hexaoleates, and combinations thereof.

3. The formulation of claim 1 or claim 2 wherein the formulation contains from about 30 % to about 48 % w/w natamycin.

4. The formulation of claim 3 wherein the formulation contains from about 35 % to about 48 % w/w natamycin.

5. The formulation of any one of claims 2 to 4 wherein the ation contains from about 0.1 % to 3.0 % w/w nonionic surfactant.

6. The formulation of any one of claims 2 to 5 wherein the nonionic surfactant is at least one kylene oxide block copolymer.

7. The formulation of any one of claims 1 to 6 comprising from about 46 % to about 75 % w/w water.

8. A stable sion trate formulation for inhibition of fungal growth comprising: about 25% w/w natamycin; from about 0.5 % to about 2.5 % w/w sodium lignosulfonate; from about 0.1 % to about 10 % w/w of a nonionic surfactant selected from the group ting of polyalkylene oxide block copolymers, polyoxyethylene sorbitan trioleates, polyoxyethylene sorbitol hexaoleates, and combinations thereof; and water, wherein the natamycin is present as particles that on average are from 3 to 9 microns in diameter and the formulation has a viscosity of less than 1400 centipoise at 21 degrees s.

9. The ation of claim 8 wherein: the sodium lignosulfonate is at a concentration of about 1.0% w/w; and the nonionic surfactant is a kylene oxide block copolymer at a concentration of about 0.5% w/w.

10. Use of the formulation of any one of claims 1 to 9 in the manufacture of a medicament for inhibiting the growth of a fungal pathogen in a t in need thereof.

11. The use of claim 10 wherein the suspension concentrate formulation further comprises from about 0.1 % to about 10 % w/w of a nonionic tant selected from the group consisting of polyalkylene oxide block copolymers, polyoxyethylene sorbitan trioleates, polyoxyethylene sorbitol hexaoleates, and combinations thereof.

12. A method of inhibiting fungal growth comprising: (a) diluting a suspension trate formulation which comprises from about 25 % to about 48 % w/w natamycin, from about 0.5% to about 2.5% w/w of sodium lignosulfonate, and water; and (b) applying the diluted suspension concentrate formulation to a commodity in need of protection from fungal growth, wherein the natamycin is present as particles that on average are from 3 to 9 microns in diameter and the ation has a ity of less than 1400 centipoise at 21 degrees Celsius prior to dilution.

13. The method of claim 12 wherein the suspension concentrate formulation r comprises from about 0.1 % to about 10 % w/w of a nonionic surfactant selected from the group consisting of kylene oxide block copolymers, polyoxyethylene sorbitan trioleates, polyoxyethylene sorbitol hexaoleates, and combinations thereof.

14. The method of claim 12 or claim 13 wherein the diluted formulation is applied by g, drenching (flooding), or spraying.

15. The method of any one of claims 12 to 14 wherein the formulation is diluted to from about 0.01 % to about 5 % w/w natamycin.

16. The method of claim 15 wherein the formulation is d to from about 0.5 % to about 2 % w/w natamycin. Valent BioSciences LLC Patent Attorneys for the Applicant/Nominated Person SPRUSON & FERGUSON