WO2007023481A1

WO2007023481A1 - Non-phytotoxic biocidal compositions for agricultural and veterinary use

Info

Publication number: WO2007023481A1
Application number: PCT/IL2005/000917
Authority: WO
Inventors: Joshua Gad-El
Original assignee: A. Shitzer Ltd.
Priority date: 2005-08-25
Filing date: 2005-08-25
Publication date: 2007-03-01

Abstract

The present invention relates to biocidal non-phytotoxic agricultural and veterinary compositions, in particular compositions effective in soil pre-planting and post planting disinfestation, as well as treatment of seeds, plants and produce comprising a mixture of at least two inorganic halides, and to the use of the compositions in agricultural applications.

Description

NON-PHYTOTOXIC BIOCIDAL COMPOSITIONS FOR AGRICULTURAL AND VETERINARY USE

FIELD OF THE INVENTION The present invention relates to non-phytotoxic biocidal agricultural and veterinary compositions, in particular compositions effective in soil pre-planting sanitation and treatment of seeds, plants and produce.

BACKGROUND OF THE INVENTION

Soil Disinfestation

Soil disinfestation is a standard procedure among crop growers in agriculture industry and in large scale home gardening. The disinfestation process is directed to the extermination of soil-borne pests such as bacteria, nematodes, fungi, viruses, protozoa, mycoplasmas and insects at different growth stages, etc., in an attempt to increase the soil's productivity and to prevent diseases from damaging or reducing agricultural yield.

Methyl bromide was the most widely used and most effective broad-spectrum soil fumigant available. In 1993, methyl bromide was listed as an ozone-depleting compound and has been phased out of use in the USA by the Clean Air Act of 1990. Alternative compositions used for soil disinfestation are not as effective as methyl bromide in eradicating a wide range of soil-borne pathogens and novel compositions are sought.

Compositions Containing Halides

Halides, such as hypochlorites (e.g. liquid sodium hypochlorite, calcium hypochlorite), trichlorocyanuric acid and n-chloro-para-toluene sulfonamide sodium salt (Chloramine^® T) are known as bleaching and sanitizing agents for domestic, industrial and to a lesser extent, agricultural use.

The halides suffer from several drawbacks. The liquid products are corrosive, unstable and readily inactivated by organic matter, thus limiting their utility and reliability, particularly on farms where large quantities of organic matter are encountered. The solid products are more stable but are much less reactive. Chloramine^® T, for example, requires extremely high concentrations to produce an acceptable biocidal effect, and exhibits reduced activity in the presence of organic matter.

EP 0 158 508 discloses an adjustable strength laundry aid package comprising two components, a bleach base composition and a bleach activator. The bleach base composition includes an oxygen-bleaching agent for use in fabric laundering while the bleach activator composition includes a halide salt or salts selected from the group consisting of chloride salts and bromide salts of an alkali metal or metals. That invention is exemplified by compositions comprising at least 50% halide salt(s). The components are packaged separately to control activation of the bleach at the time of fabric laundering.

ZA 9701266 discloses a composition for the sanitation of water comprising a disinfecting component selected from sodium bromide, potassium bromide, ammonium bromide, potassium iodide, sodium iodide, trichloroisocyanurate, sodium chloride or a combination thereof, and an activator component, the activator component being an oxidizing agent. There is no exemplification of a composition comprising a mixture of inorganic halides

U.S. Patent No. 3,337,466 ("the '466 patent") discloses effervescent compositions for cleaning dentures comprising a mixture of from about 5 to about 40 parts of a mixture of potassium monopersulfate, potassium sulfate and potassium hydrogen sulfate (Oxone®); from about 40 to about 5 parts of an inorganic water soluble peroxide of a Group I or II metal; up to about 5 parts of a water soluble halide selected from chloride, bromide and iodide of an alkali or alkaline earth metal and of ammonium; from about 15 to about 85 parts of a water-soluble inorganic alkaline filler; a chelating agent and a wetting agent. The '466 patent discloses that an aqueous solution derived from the composition has a pH of least 7. A composition comprising a plurality of water- soluble halides is neither taught nor suggested in that patent.

U.S. Patent No. 4,822,512 ("the '512 patent") discloses disinfecting, particularly virucidal, compositions comprising 0.01 to 5 parts by weight of a total of 100 parts of an inorganic halide, preferably sodium chloride; an oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions; sulfamic acid; an optional non-reducing organic acid and an anhydrous alkali metal phosphate. The '512 patent demonstrates the use of a composition comprising sodium chloride for veterinarian applications and water sanitation, for example in livestock and fishery farms. The '512 patent neither teaches nor suggests a composition comprising a plurality of inorganic halides nor the use of the compositions in soil sanitization or seed disinfection. There exists a need for improved halide-based biocidal compositions for use in agricultural applications. Specifically, the need exists for non-phytotoxic formulations effective for soil disinfestation both prior to seeding and during cultivation.

SUMMARY OF THE INVENTION

The present invention is directed to novel non-phytotoxic biocidal/antimicrobial compositions and methods of use thereof. The present invention is based on the unexpected discovery that a composition comprising a mixture of two or more inorganic halides possesses increased biocidal activity when compared to a composition comprising a single inorganic halide. The biocidal compositions of the invention comprise novel combinations of known ingredients that show an unexpected synergistic effect compared to hitherto known compositions.

The compositions have utility in agricultural applications, in particular in agronomic and veterinary applications.

The components of the present composition are provided as percent weight per total weight (% wt/wt). In one aspect, the present invention provides a dry, water-soluble, non-phytotoxic biocidal composition comprising:

(a) from about 3% wt/wt to about 20% wt/wt of a water-soluble mixture of at least two inorganic halides;

(b) at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions;

(c) sulphamic acid;

(d) at least one non-reducing organic acid;

(e) at least one anhydrous alkali metal phosphate; (f) at least one excipient selected from the group consisting of nonorganic inert chemical filler, surfactant and a combination thereof; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5.

The above composition, comprising a mixture of two or more inorganic halides, is now shown to produce an unexpected synergistic biocidal effect in eradicating pathogens. This effect is evidenced in experiments comparing the composition of the present invention to a similar composition comprising a single halide. In one embodiment the present invention provides a composition comprising:

(b) from about 25% wt/wt to about 60% wt/wt of at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions;

(c) from about 3% wt/wt to about 10% wt/wt sulphamic acid;

(d) from about 3% wt/wt to about 20% wt/wt of at least one non- reducing organic acid;

(e) from about 5% wt/wt to about 30% wt/wt of at least one anhydrous alkali metal phosphate;

(f) from about 1% wt/wt to about 61% wt/wt of non-organic inert chemical filler; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5. In some embodiments the composition further comprises a surfactant. The presence of one or more surfactants in the composition may be preferred in certain applications.

In another embodiment the present invention provides a composition comprising:

(a) from about 3% wt/wt to about 20% wt/wt of a water-soluble mixture of at least two inorganic halides; (b) from about 25% wt/wt to about 60% wt/wt of at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions;

(c) from about 3% wt/wt to about 10% wt/wt sulphamic acid; (d) from about 3% wt/wt to about 20% wt/wt of at least one non- reducing organic acid;

(f) from about 5% wt/wt to about 20% wt/wt of a surfactant; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5.

In some embodiments the inorganic halide mixture compromises a mixture of inorganic halides selected from metal halides and ammonium halides. Typical metal halides include alkaline earth and alkali metals. Preferred are sodium and potassium alkali metals. In some embodiments the inorganic halide mixture comprises two or more metal halides selected from the group consisting of potassium chloride, potassium bromide, potassium iodide, sodium bromide, sodium chloride and sodium iodide. Fluoride compounds may be toxic to plants and animals.

In preferred embodiments the inorganic halide mixture is a mixture of a chloride salt and a bromide salt. In specific embodiments the inorganic halide mixture is a potassium chloride and sodium bromide mixture. In certain embodiments the inorganic halide mixture is a sodium chloride and sodium bromide mixture. In other embodiments a mixture of potassium chloride and potassium bromide or a mixture of sodium chloride and potassium bromide is provided. In specific embodiments the inorganic halide mixture excludes an iodide salt.

According to the present invention the inorganic halide mixture is provided at a range of from about 3% wt/wt to about 20% wt/wt. In some embodiments the inorganic halide mixture is provided at a range of from about 5% wt/wt to about 15% wt/wt. In other embodiments the inorganic halide mixture is provided at about 10% w/w. The inorganic halides can be provided in various proportions. In some embodiments the ratio of bromide salt to chloride salt is about 3 :7 wt/wt to about 9: 1 wt/wt. In preferred embodiments the bromide salt to chloride salt are provided at about 1 : 1 ratio wt/wt. In preferred embodiments the inorganic halide mixture comprises about 5% wt/wt bromide salt and about 5% wt/wt chloride salt.

According to the present invention at least one oxidizing agent is preferably selected from an agent capable of reacting with an inorganic halide to generate hypohalite ions. The at least one oxidizing agent is provided in an amount from about 25% wt/wt to about 60% wt/wt. In some embodiments at least one oxidizing agent is provided in an amount about 35% wt/wt to about 55% wt/wt. In specific embodiments at least one oxidizing agent is provided at about 50% w/w. In some embodiments the oxidizing agent may be selected from the group consisting of a persulfate compound and a peroxyphthalate compound. In certain embodiments the oxidizing agent is a peroxyphthalate compound e.g., potassium monoperoxyphthalate. In certain preferred embodiments the oxidizing agent is a persulfate compound. In specific embodiments the persulfate compound is potassium hydrogen peroxymonosulfate sulfate.

Sulfamic acid is an provided in an amount of about 3% wt/wt to about 10% wt/wt. In some embodiments, sulfamic acid is provided at about 5% wt/wt to about 8% wt/wt. In a currently preferred embodiment sulfamic acid is provided at about 5% wt/wt.

At least one non-reducing organic acid is provided in the present composition in an amount of about 3% wt/wt to about 20% wt/wt. In some embodiments the non- reducing organic acid is provided in an amount about 5% wt/wt to about 15% wt/wt. In specific embodiments a non-reducing organic acid is provided in an amount of about 8% wt/wt to about 10% wt/wt.

In one embodiment the organic-based acid is selected from malic acid, tartaric acid, citric acid, acetic acid, glycolic, glutamic acid, sorbic acid, benzoic acid, or dimeric acids such as oxalic acid, succinic acid, glutaric acid, adipic acid, maleic acid, fumaric acid, etc., or fatty acids such as butyric acid, caprylic acid, caproic acid, octanoic acid, nonanoic acid, decanoic acid, and the like; or mixtures thereof. In some embodiments the at least one non-reducing organic acid is selected from malic acid, adipic acid and succinic acid. In specific embodiments the non-reducing organic acid is malic acid.

In one embodiment, at least one alkali metal phosphate is provided in the present composition in an amount of about 5% wt/wt to about 30% wt/wt. In some embodiments an alkali metal phosphate is provided at about 10% wt/wt to about 20% wt/wt. In a currently preferred embodiment an alkali metal phosphate is provided at about 10% wt/wt.

The anhydrous alkali metal phosphate can be any alkali metal phosphate known in the art. In some embodiments the anhydrous alkali metal phosphate is selected from sodium hexametaphosphate, tetrasodium diphosphate, tetrasodium pyrophosphate, mono- sodium phosphate, di-sodium phosphate, tri-sodium phosphate, mono-potassium phosphate (MKP), di-potassium phosphate, tri-potassium phosphate and combinations thereof. According to a currently preferred embodiment, the alkali metal phosphate is MKP. In one preferred embodiment MKP is provided in an amount of about 10% wt/wt of the composition.

In one embodiment, chemical filler is an inert inorganic hygroscopic salt selected from the group consisting of calcium carbonate, magnesium silicate, sodium silicate, sodium carbonate, calcium bicarbonate, potassium bicarbonate and combinations thereof. In certain embodiments, the chemical filler is calcium carbonate.

A surfactant compatible with the acids and oxidizing agents is acceptable. The surfactant may be selected from the group consisting of lauryl ether sulfates, ethylene oxide, propylene oxide aklyl phenol condensates, polyglycol ethers of fatty alcohols, fatty acid ethylene oxide condensates, polyglycol ethers of alkyl phenols, fatty alcohol ethoxylates and combinations thereof. A preferred surfactant is sodium dodecylbenzene sulphonate. The surfactant can be provided in the composition at a concentration of about 5% wt/wt to about 20% wt/wt. Preferably, the surfactant is provided in an amount of about 10% wt/wt.

In one preferred embodiment the present invention provides a composition comprising:

(a) about 10% wt/wt of a water-soluble mixture of potassium chloride and sodium bromide in equal amounts wt/wt;

(b) about 50% wt/wt of potassium hydrogen peroxymonosulfate;

(c) about 5% wt/wt sulphamic acid; (d) about 8% wt/wt of malic acid;

(e) about 10% wt/wt of monopotassium phosphate; (f) about 17% wt/wt of at least one non-organic inert chemical filler; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being about 2.4. The composition is preferably prepared as a solid in a form selected from the group consisting of powder, tablet, dispersion, granules, pill, capsule and microcapsules. The compositions may be prepared as controlled-release or slow-release compositions. Preferably the composition is provided as a powder that is soluble in an aqueous medium. In some embodiments the powder is applied as is. In certain embodiments an aqueous solution is prepared immediately prior to use. In some embodiments the composition is prepared as a semi-solid, for example as a paste or cream.

In another aspect methods of using the compositions are provided. In some embodiments the compositions are useful as a soil sanitizer. The compositions are non- phytotoxic and therefore show utility in pre-planting soil sterilization and post-planting soil treatment.

To facilitate the distribution of the composition on large tracts of land, a dilution of the composition into a solid, inert compound may be desired. For certain applications, the composition of the present invention is diluted about 2- to about 7-fold with nonorganic, inert chemical filler. In certain preferred embodiments the composition is diluted about 4-fold.

Accordingly, the present invention further provides a non-phytotoxic biocidal composition for soil disinfestation comprising about 15% wt/wt to about 50% wt/wt of a composition comprising

(a) from about 3% wt/wt to about 20% wt/wt of a water- soluble mixture of at least two inorganic halides;

(c) from about 3% wt/wt to about 10% wt/wt sulphamic acid; (d) from about 3% wt/wt to about 20% wt/wt of at least one non-reducing organic acid; (e) from about 5% wt/wt to about 30% wt/wt of at least one anhydrous alkali metal phosphate;

(f) at least one non-organic inert filler the pH of a 1% by weight aqueous solution of the composition being between 1.2 and 5.5; and about 50% wt/wt to about 85% wt/wt of a non-organic, inert chemical filler.

In some embodiments the non-organic chemical filler is selected from the group consisting of calcium carbonate and sodium carbonate (soda ash).

The method of applying the composition is not limiting. In some embodiments the composition is nsedper se. In other embodiments the composition is blended with non-organic inert chemical filler. In one embodiment the present invention provides a method for soil disinfestation comprising the step of: a) applying to the soil a biocidal-effective amount a composition according to the present invention. In another aspect the method further comprises the step of: b) blending said composition into the soil.

For soil pre-planting sanitation, it is possible to apply a solid composition, including powder, to the surface of the soil and optionally blend it into the soil to a predetermined depth by methods known in the art, including tilling and cultivation. Application of the powder in this manner will ensure that the biocidal activity is present throughout the soil layers. In some embodiments the composition is formulated as liquid and can be applied using suitable means, including by spray or drip irrigation.

Furthermore, the compositions of the present invention are useful in seed and fish disinfection. Accordingly, the present invention provides a method for the disinfection of fish comprising the steps of: a) preparing a solution comprising a biocidally-effective amount of a composition comprising:

(i) from about 3% wt/wt to about 20% wt/wt of a water-soluble mixture of at least two inorganic halides; (ii) at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions;

(iii) sulphamic acid;

(iv) at least one non-reducing organic acid; (v) at least one anhydrous alkali metal phosphate;

(vi) at least one excipient selected from the group consisting of non-organic inert chemical filler, surfactant and a combination thereof; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5; b) exposing fish to said solution.

The solution may further comprise about 1% wt/v to about 5% wt/v pyrogen.

The solution is effective in eradicating parasites and microbes that infect fish. A biocidally effective solution comprises about 0.05% wt/v to about 20% wt/v of the composition, preferably about 0.5% wt/v to about 2% wt/v of the composition in order to be an effective, yet harmless, disinfectant of the fish. The composition is effective in treating fish tanks, aquariums and is useful in treating fishponds and fish farms.

The present invention also provides a method for the disinfection of seeds comprising the steps of: a) preparing a solution comprising a biocidally-effective amount of a composition comprising:

(i) from about 3% to about 20% of a water-soluble mixture of at least two inorganic halides;

(ii) at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions; (iii) sulphamic acid;

(iv) at least one non-reducing organic acid;

(v) at least one anhydrous alkali metal phosphate; (vi) at least one excipient selected from the group consisting of non-organic inert chemical filler, surfactant and a combination thereof; and; wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5; and b) exposing the seeds said solution for sufficient time to disinfect said seeds.

The solution is effective in disinfecting seeds prior to planting. A biocidally effective solution comprises about 0.05% wt/v (weight per volume) to about 20% wt/v composition, in order to be effective in disinfection, yet harmless to the seeds. In some embodiments a seed disinfecting solution comprises about 0.1% wt/v to about 5% wt/v of the composition of the present invention. Sufficient time for disinfection is preferably less than 1 hour, and more preferably less than 30 minutes.

The composition of the present invention is also useful in treating post-harvest produce and in disinfecting animal and their surroundings. These and other embodiments of the present invention will become apparent in conjunction with the figures, description and claims that follow.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 demonstrates the level of CFU (colony forming units) of microorganisms present in the soil following treatment of soil with the compositions of the present invention.

Figure 2 shows seedlings sprouted from seeds treated with composition 21 and composition 22.

Figure 3 demonstrates the level of disinfection offish as a reduction in CFU following treatment with composition 21. DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to novel antimicrobial/disinfestation compositions and utility thereof, particularly in agricultural applications.

The invention discloses for the first time a composition comprising a plurality of inorganic halides as a highly effective disinfectant. In particular, the composition is an effective, non-phytotoxic antimicrobial agent suitable for soil disinfestation, seed disinfestation, and animal disinfection. The composition is applicable as a soil sanitation composition both for use prior to planting and during the growing season.

Advantages of Present Invention The advantages of the present composition include the following: a) The compositions comprise a combination of at least two inorganic halides, which provide an unexpectedly effective biocidal activity; b) Non-phytotoxic composition that is highly effective for soil disinfestation before seeding or planting and for soil treatment during the growing season; c) Broad spectrum composition effective in eradication soil pests including plant root viral, nematode and bacterial diseases; d) Safe for use on and within vicinity of animals, including fish and livestock; e) The reactivation process results in reduction in the total amount of halogen needed for the sterilization process, thereby reducing the soil and water source contamination. f) The compositions are stable and active in soil and in the presence of organic matter; g) The compositions are stable and do not liberate halide gas in the dry state.

Halides are released upon exposure to water at a pH less than 3. h) The compositions can be applied to the soil as a liquid composition or as a dry composition; i) Composition comprising a low bromide concentration, compared to methyl bromide soil fumigation, thereby eliminating danger of ozone damage; j) The danger associated with chlorine evolution is obviated while enhancing the biocidal activity through the use of a mixture of two or more halides.

Without wishing to be bound to theory, the phosphates act as buffering and chelating agents, in combination with the pH stability of sulphamic acid, enable the compositions of the present invention to be effective over a wide range of conditions. For example, the composition can be dissolved in hard water or in seawater without deleteriously affecting its biocidal properties.

Preferred Embodiments

According to a first aspect, the invention provides a dry, water-soluble biocidal composition comprising (a) from about 3% to about 20% wt/wt of a water-soluble mixture of at least two inorganic halides;

(c) sulphamic acid; (d) at least one non-reducing organic acid;

(e) at least one anhydrous alkali metal phosphate;

(f) at least one excipient selected from the group consisting of nonorganic inert chemical filler, surfactant and a combination thereof; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5.

In one embodiment, the present invention provides a dry, water-soluble biocidal composition comprising: (a) from about 3% to about 20% wt/wt of a water-soluble mixture of at least two inorganic halides;

(b) from about 25% to about 60% wt/wt of at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions;

(c) from about 3% to about 10% wt/wt sulphamic acid;

(d) from about 3% to about 20% wt/wt of at least one non- reducing organic acid;

(e) from about 5% to about 30% wt/wt of at least one anhydrous alkali metal pho sphate;

(g) from about 1% to about 61% wt/wt of non-organic inert chemical filler; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5. In some embodiments the composition further comprises from about 1% to about

61% wt/wt of at least one non-organic inert chemical filler.

In one embodiment, the present invention provides a dry, water-soluble biocidal composition comprising:

(a) about 5% to about 15% wt/wt of a water-soluble mixture of at least two inorganic halides;

(b) about 35% to about 55% wt/wt of at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions;

(c) about 5% to about 8% wt/wt sulphamic acid; (d) from about 5% to about 15% wt/wt of at least one non- reducing organic acid;

(e) from about 10% to about 20% wt/wt of at least one anhydrous alkali metal phosphate;

(f) from about 10% to about 40% wt/wt of non-organic inert chemical filler; the pH of a 1% by weight aqueous solution of the composition being between 1.2 and 5.5. In some embodiments the combination of non-organic inert filler and a surfactant is included in the composition. In some embodiments a composition devoid of nonorganic inert filler is preferred. Accordingly, the present invention provides a dry, non- phytotoxic composition comprising: (a) from about 3% to about 20% wt/wt of a water-soluble mixture of at least two inorganic halides;

(b) from about 25% to about 60% wt/wt of at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions; (c) from about 3% to about 10% wt/wt sulphamic acid;

(d) from about 3% to about 20% wt/wt of at least one non-reducing organic acid;

(e) from about 5% to about 30% wt/wt of at least one anhydrous alkali metal phosphate; (f) from about 5% to about 20% of surfactant; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5.

The term "dry" refers to a composition comprising less than 10% moisture, and preferably less than 5% moisture.

The term "biocidal" refers to the reduction in the number of one or more pathogen species in a treated sample compared to an untreated sample. In one embodiment a reduction of at least 10% in the number of at least one pathogen is biocidally effective. In preferred embodiments, a reduction of at least 50% is biocidally effective.

In one embodiment, the inorganic halides comprise a mixture of two water- soluble inorganic halides. In certain embodiments the inorganic halides are provided in equal amounts wt/wt. In some embodiments the inorganic halide mixture compromises a mixture of inorganic halides selected from metal halides and ammonium halides. Typical metal halides include alkaline earth and alkali metals. Preferred are sodium and potassium alkali metals. In one preferred embodiment, the inorganic halide is a mixture of potassium chloride and sodium bromide. In certain embodiments the inorganic halide mixture is provided at a concentration of about 3% to about 20% w/w. In other embodiments the inorganic halide mixture is provided at a concentration of about 5% to about 15% w/w. In other embodiments the inorganic halide mixture is provided at about 10 % wt/wt.

Although the preferred inorganic halide mixture is potassium chloride and sodium bromide mixture, any other mixture of inorganic halides can be substituted, for example, mixtures of potassium chloride, potassium bromide, potassium iodide, sodium bromide, sodium chloride or sodium iodide, are acceptable provided that they do not react with the components of the composition to form an insoluble salt. In some embodiments the inorganic halide mixture is a mixture of potassium chloride and potassium bromide. In other embodiments the inorganic halide mixture is a mixture of sodium chloride and sodium bromide.

The present composition further contemplates compositions comprising an inorganic halide mixture comprising a mixture of three or more inorganic halides. In some embodiments, the composition comprises, for example, a mixture of NaCl, KCl and KBr.

In general, as the inorganic halide concentration is increased in the composition, the amount of non-organic inert chemical filler is increased proportionately. Without wishing to be bound to theory, the filler acts as a buffer.

In some embodiments at least one oxidizing agent is provided in an amount about 35% to about 55% wt/wt. In specific embodiments the at least one oxidizing agent is provided at about 50% wt/wt.

The oxidizing agent is preferably selected from an agent capable of reacting with the inorganic halides to generate hypohalite ions. In some embodiments the oxidizing agent can be selected from the group consisting of a persulfate compound and a peroxyphthalate compound. In certain embodiments the oxidizing agent is a peroxyphthalate compound and in particular embodiments is potassium monoperoxyphthalate. In certain preferred embodiments the oxidizing agent is a persulfate compound. In specific embodiments the persulfate compound is the commercially available potassium hydrogen peroxymonosulfate (Oxone®), represented by the chemical formula 2KHSO₅-KHSO₄-K₂SO₄ [CAS-RN 70693-62-8]. Sulfamic acid is provided in an amount of about 3% to about 10% wt/wt. In some embodiments, sulfamic acid is provided at about 5% to about 8% wt/wt. In some embodiments, sulfamic acid is provided at about 5% wt/wt.

At least one non-reducing organic acid is provided in the present composition in an amount of about 3% to about 20% wt/wt. In some embodiments the non-reducing organic acid is provided in an amount of about 5% to about 15% wt/wt. In certain embodiments the non-reducing organic acid is provided in an amount of about 5% to about 10% wt/wt. In specific embodiments a non-reducing organic acid is provided in an amount of about 8% wt/wt. In some embodiments the at least one non-reducing organic acid is selected from malic acid, adipic acid and succinic acid. In specific embodiments the non-reducing organic acid is malic acid.

At least one alkali metal phosphate is provided in the present composition in an amount of about 5% to about 30% wt/wt. In some embodiments an alkali metal phosphate is provided at about 10% to about 20% wt/wt.

The alkali metal phosphate can be any alkali metal phosphate known in the art. In some embodiments the alkali metal phosphate is selected from sodium hexametaphosphate, tetrasodium pyrophosphate, mono- sodium phosphate, di-sodium phosphate, tri-sodium phosphate, mono-potassium phosphate (MKP), di-potassium phosphate, tri-potassium phosphate and mixtures thereof.

In some embodiments the composition comprises about 10% MKP as a preferred alkali metal phosphate.

In one preferred embodiment, the invention provides a dry, water-soluble biocidal composition comprising: (a) about 10% wt/wt mixture of potassium chloride and sodium bromide

(b) about 50% potassium hydrogen peroxymonosulfate;

(c) about 5% wt/wt sulphamic acid;

(d) about 8% wt/wt malic acid; (e) about 10% wt/wt MKP;

(f) about 17% wt/wt of calcium carbonate; and the pH of a 1% by weight aqueous solution prepared from said composition being about 2.4.

In certain preferred embodiments the inorganic halide mixture comprises potassium chloride (KCl) and sodium bromide (NaBr). This composition was unexpectedly found to be substantially more effective than hitherto known compositions comprising a single halide. As is demonstrated in the Examples hereinbelow, the composition of the invention was unexpectedly found to be about 100 times more active as a soil-sanitizing agent than a composition comprising a single halide, specifically a composition comprising sodium chloride composition 22 or Virkon^®-S. In some embodiments the inorganic halide mixture comprises sodium chloride

(NaCl) and sodium bromide (NaBr). In yet other embodiments the inorganic halide mixture comprises potassium chloride (KCl) and potassium bromide (KBr).

Without wishing to be bound to theory, the composition of the present invention is able to reactivate the halides, resulting in reduced halide consumption. Thus, the biocidal advantages of bromides, and the specific synergistic effect of the bromide- chloride formulation, can be utilized with minimized environmental damage.

The presence of one or more surfactants in the composition is optional and can be preferred in some applications, in particular for seed disinfection and veterinary applications. A surfactant compatible with the acids and oxidizing agents is acceptable. The surfactant can be selected from the group consisting of lauryl ether sulfates, ethylene oxide aklyl phenol condensates, propylene oxide aklyl phenol condensates, polyglycol ethers of fatty alcohols, fatty acid ethylene oxide condensates, polyglycol ethers of alkyl phenols, fatty alcohol ethoxylates and combinations thereof. Preferred surfactants include sodium dodecylbenzene sulphonate and sodium lauryl benzene sulfate. The surfactant can be provided in the composition at a concentration of about 5% wt/wt to about 20% wt/wt.. Preferably the surfactant is provided to about 10% wt/wt.

Utility

The present invention further provides utility and methods of using the disclosed compositions. The compositions of the invention are useful in diverse applications due to their low toxicity to plants and animals. The compositions of the present invention have been shown to be effective in soil disinfestation for the effective control of soil-borne plant pathogens, including soil-borne pests such as bacteria, nematodes, fungi, viruses, protozoa, mycoplasmas and insects. The compositions are useful in the treatment of soil either before planting or during the growing season. Plant pathogens cause significant global agricultural devastation annually, including damage to and loss of crops and flowers in nurseries, greenhouses, fields and groves.

Bacterial pathogens have a significant impact. Bacterial pathogens include species of Pseudomonas; Erwinia; Agrobacteria; Xanthomonas; Clavibacter; Xylella and others. For example, Erwinia species cause blight, wilt, or soft-rot in numerous trees, flowers, produce and crops. Xanthomonas species cause spots, wilts, rots and cankers in a variety of citrus trees and vegetable plants.

Fungal plant pathogens include species from the genera Alternaria; Ascochyta; Botrytis; Cercospora; Colletotrichum; Diplodia; Erysiphe; Fusarium; Gaeumanomyces; Helminthosporium; Macrophomina; Nectria; Peronospora; Phoma; Phymatotrichum; Phytophthora; Plasmopara; Podosphaera; Puccinia; Pythium; Pyrenophora; Pyricularia; Rhizoctonia; Scerotium; Sclerotinia; Septoria; Thielaviopsis; Uncinula; Venturia; Verticillium and others.

Plant pathogenic nematodes include nematodes of the economically important Heterodera; Meloidogyne and Pratylenchus species among others causing diseases such as root galls, root rot, stunting and various other rots. In addition, nematodes serve as vectors for viral plant pathogens.

Economically significant viral plant pathogens include members of the Potyviridae family of viruses. Agriculturally important insects include those species that attack plants directly and those that are vectors of plant pathogenic microbes.

The method of applying the compositions is not limiting. The compositions can be added to the soil in solid, semi-solid or liquid form. As a solid the composition can be added to the soil in an amount of about 100 kg to about 500 kg per 1,000 square meters (m²) of a 1:4 diluted composition. As a liquid about 250 liter (1) to about 1000 1 of a 10% solution is adequate. Depending on the type of soil and depth of application more or less material can be added to the soil. In some instances, a dilution of the solid composition is preferred for application to the soil. Accordingly, the present invention further provides a solid composition for soil disinfestation comprising about 15% wt/wt to about 50% wt/wt of a composition comprising (a) from about 3% wt/wt to about 20% wt/wt of a water- soluble mixture of at least two inorganic halides; (b) from about 25% wt/wt to about 60% wt/wt of at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions; (c) from about 3% wt/wt to about 10% wt/wt sulphamic acid;

(d) from about 3% wt/wt to about 20% wt/wt of at least one non-reducing organic acid;

(e) from about 5% wt/wt to about 30% wt/wt of at least one anhydrous alkali metal phosphate; (f) from about 1% wt/wt to about 61% wt/wt of at least one non-organic inert chemical filler; the pH of a 1% by weight aqueous solution of the composition being between 1.2 and 5.5; and about 50% wt/wt to about 85% wt/wt of a non-organic, inert chemical filler.

In one embodiment, the method for soil disinfestation comprises the step of a) applying to the soil a biocidal-effective amount of the composition of the present invention. In additional embodiments the method for soil disinfestation further comprises the step of: b) blending said composition into the soil.

Additionally, the compositions of the present invention show utility in seed disinfection and in fish farm ponds and aquarium disinfection.

Accordingly the present invention provides a method for seed disinfection comprising the steps of: a) preparing a dilute aqueous solution of the composition of the invention; and , b) wetting the seeds in said solution. A typical seed treatment encompasses immersing seeds in a 0.05% to about 20% dilute solution of the composition. Preferably the seeds are treated in a 0.1% to about 5% dilute solution for about 5 minutes to about 60 minutes.

Furthermore, the present invention provides a method for the disinfection offish comprising the steps of: a) preparing a dilute aqueous solution of the composition of the invention; b) exposing fish to said solution.

In some embodiments the composition is added to a vessel containing fish including an aquarium, bowl, pond, lake, pool, fish farm and the like.

The following examples are presented in order to more fully illustrate some embodiments of the invention. They should, in no way be construed, however, as limiting the broad scope of the invention.

EXAMPLES

Example 1 : Preparation of a Composition

A biocidal composition, referred to herein composition 21, was prepared by mixing together the following ingredients:

In addition, a similar composition comprising a single halide, specifically 10% NaCl, was prepared and used for comparison to composition 21 in different applications. The composition comprising a single halide is referred to herein composition 22.

The potassium persulphate triple salt (potassium hydrogen peroxymonosulfate) has the general formula 2KHSO₅*KHSO₄-K₂SO₄ and is commercially available as Oxone®. The mono potassium phosphate, known as MKP, is provided as powder or granules.

Composition 21 was prepared by first preparing a potassium chloride/sodium bromide mixture by mixing together equal amounts by weight of the two salts. The potassium chloride/sodium bromide mixture was blended with the chemical filler, calcium carbonate. The compounds MKP, sulphamic acid, malic acid were added to the halide-filler mixture and finally, the resulting mixture was blended with the persulfate. A surfactant is optionally added. Composition 21 is stable and has a long shelflife.

A 1% by weight solution of the composition in de-ionized water had a pH of about 2.4. The pH of the solution may vary, depending on the water source.

For soil disinfestation of large areas, including fields, pastures and greenhouses, it may be desirable to use a dilute solid composition comprising a mixture of at least two inorganic halides, which is easily distributed and can be more uniformly dispersed through the soil. Therefore, the above composition was diluted (dry dilution) about 2-to about 7-fold in dry inert chemical filler, and tested for its biocidal activity in soil. A fourfold diluted composition is preferred.

Virkon^®-S is a commercially available (Antec, division of DuPont) biocide useful in cleaning and disinfection. Virkon^®-S comprises a single halide (sodium chloride), potassium peroxomonosulfate, sulfamic acid, malic acid, sodium hexametaphosphate, and 15% surfactant.

Example 2: Eradication of soil-borne pathogens

Soil borne pathogens, including bacteria, nematodes, fungi, viruses, insects and mycoplasmas provide a significant source of infection and economic damage to crops and ornamentals. Furthermore, the pathogens persist in the soil following plant removal and need to be eradicated prior to the subsequent planting season. Often, pathogens will infect plants and/or soil during the growth season and require prompt eradication.

Fungi

Tests have been carried out to establish the biocidal activity of the above composition in accordance with the standard in soil pre-planting sanitation and disinfecting treatments of seeds. Pathogenic fungi, such as Fusarium species are both seed-borne and soil-borne. These pathogens affect many different hosts, including crops and ornamentals and result in immeasurable economic damage. Fusarium spp. is difficult to eradicate since compositions known to treat Fusarium are often toxic to the hosts.

The biocidal activity of the composition was tested by applying the present composition onto the soil surface and incorporating into the soil to various depths by tilling and or cultivation.

Composition 21, was compared to composition 22 and to a commercially available composition, Virkon^®-S for efficacy in eradication of soil-borne pathogens.

Soil was infected with either a mixture of soil-borne pathogens including bacteria or Fusarium fungi. Columns were filled with infected soil. Compositions 21, 22 or Virkon^®-S were added to the column. The soil was watered and the soil was tested for pathogens at different depths.

At a depth of 5 cm the three compositions had approximately equal activity. Table 1 shows the number of colony forming units (CFU) of Fusarium remaining in the soil at a depth of 5 cm. Control refers to untreated soil. All tests were performed in triplicate. All Pairwise Multiple Comparison Procedures (Fisher LSD Method) were used to compare the different treatments to each other. A statistically significant results were obtained for all tests comparing controls to composition 21.

Table 1 : Fusarium remaining in treated soil at a depth of 5 cm.

At a depth of 25 cm (Table 2) the biocidal activity of the composition 21 became apparent. While more than 50% of the fungi remained following treatment with Virkon^®- S, and greater than 90% of the fungi remained following treatment with composition 22 having a single halide, no fungi were detected in the soil treated with composition 21.

The above results are presented as a graph in Figure 1. Table 2: Fusarium remaining in treated soil at a depth of 25 cm.

Normality Test: Passed (P > 0.200); Equal Variance Test: Passed (P = 0.221)

The differences in the mean values among the treatment groups are greater than would be expected by chance; there is a statistically significant difference (P = <0.001). To isolate the group or groups that differ from the others a multiple comparison procedure was used. Power of performed test with alpha = 0.050: 1.000.

At a depth of 39 cm the differences in the biocidal activity between the composition of the present invention and the composition comprising a single halide (22 and Virkon^®-S) are significant. Table 3 shows that composition 21 eradicated all the fungi while composition 22- and Virkon^®-S-treated soil still has high numbers of fungi.

Table 3: Fusarium remaining in treated soil at a depth of 39 cm.

Normality Test: Passed (P > 0.200); Equal Variance Test:Passed (P = 0.362)

Bacteria

The compositions of the present invention are useful soil disinfestation of bacteria. Soil-borne pathogens were added to the column and the number of bacteria (CFU) was counted. At a depth of 5 cm composition 21 was at least 10 times more effective and up to about 50 times more effective in eradicating bacteria (see table 4)

Table 4: Bacteria remaining in treated soil at a depth of 5 cm.

Pairwise multiple comparison procedures were performed using the Student- Newman-Keuls method. All comparisons were statistically significant (P< 0.05)

Table 5: Bacteria remaining in treated soil at a depth of 25 cm.

SOIL Control 21 22 Virkon^®-S DEPTH

25 10.0 x 10^y 4. O x 10⁶ 7 .O x 10^b 3.O x IO⁷

25 7.0 x l0^y 3. 6 x IO³ 8 .O x 10⁶ 4.O x IO⁷

25 8.7 x lO^y 3. 6 x IO³ 6 .4 x 10^b 2.9 x lO⁷

Table 6: Bacteria remaining in treated soil at a depth of 39 cm.

Conclusions: In all sets of experiments, composition 21 performed significantly better that the commercially available mono-halide composition Virkon^®-S or composition 22. Composition 21 was able to eradicate large numbers of pathogens in the soil, at various depths at about 10 to about 100 fold better than the monohalide compositions.

Example 4: Eradication of the bacteria, Erwinia carotavora, in Ivy plants

Composition 21 was tested for effectiveness in treating Erwinia carotovora infection of Small leaf Ivy (Hedera helix). The composition was effective in eradicating the bacteria. Dilute aqueous solutions of 0.1%, 0.2%, and 0.3% of composition 21 were prepared. The product was applied to a 1 square meter quadrant at a rate of 1 liter of solution per meter by spray. The trial was repeated twice at each rate. No phytotoxicity signs have appeared after 48 hours the spread of the bacteria was arrested by the treatment. Example 5: Disinfection of Cauliflower Seeds

A composition further comprising a surfactant, sodium dodecylbenzene sulphonate, was added to the composition 21 to a final concentration of 10%. This composition is referred to herein as composition 23.

Cauliflowers seeds were infested with a mixture of soil-borne pathogens. The number of total bacterial, Xanthomonas, total fungi and Aspergillus was counted before and following treatment. Seeds were immersed in solutions of composition 23, diluted to a 0.1%, 0.5%, 1% or 2% aqueous solution in distilled water. Solutions of about 0.05% to about 20% w/v are acceptable. Table 7 shows the results of the experiment measured as CFU.

Table 7. Number of bacteria and J fungi remaining on seeds following treatment with aqueous solution of composition 23

Concentration of Time # Total # # Total # composition 23 (min.) bacteria Xanthomonas fungi Aspergillus untreated 10 3.O x IO⁵ 1.O x 10^J 2.4 x lO² 1.0 x lO^'2 untreated 20 3.I x IO³ 2.O x 10^d 2.3 x 10² 1.0 x lO²

0.1% 10 7.0 x l0⁴ 2.O x 10^J 1.7 x lO² LO x IO¹

0.1% 20 9.0 x lO" 3.O x 10² 9.O x IO¹ 0.0

0.5% 10 6.0 x 10" 1.O x 10¹ 0.0 0.0

0.5% 20 1.0 x 10⁴ 0.0 4.O x IO¹ 0.0

1.0% 10 3.O x IO² 1.O x 10¹ 0.0 0.0

1.0% 20 1.3 x l0^J 0.0 0.0 0.0

2.0% 10 1.0 x 10¹ 0.0 0.0 0.0

2.0% 20 0.0 0.0 0.0 0.0

The results show that an aqueous solution of 2% of composition 23 eradicates most of the bacteria infecting the seeds, and all of the Xanthomonas species. An aqueous solution of 0.5% of composition 23 eradicates more than 99% of the total fungi and all of the Fusarium species.

Figures 2A-C shows sprouting seeds following treatment with composition 23 compared to untreated seeds. The petri dish on the left in Figure 2A shows untreated seeds that have begun to send out roots. Even at such an early stage, the start of fungal infection can be seen (marked by arrows), whereas the treated seeds in the petri dish on the right show no signs of fungal growth. Figure 2B shows seedling grown from untreated seeds that have been stunted by the fungal growth (arrows). Figure 2C shows that seedlings grown from seeds treated with composition 23 are disease free.

Example 6: Disinfection of Fish

Fish were disinfected by exposing them to 0.1% aqueous solution of composition 24, a composition comprising a 10% mixture of KCl and NaBr and 10% surfactant (sodium dodecylbenzenesulphonate) in place of chemical filler, and optionally about about 1% to about 5% pyrogen. An aqueous solution comprising about 0.05% to about 20% wt/v of composition 24 is acceptable.

Table 8 shows the reduction in the percent of infected fish over time following treatment with composition 21, composition 24 or malachite green (M. G.).

Table 8: Percent of Infected Fish

Example 7: Treatment of Livestock

Livestock are sensitive to infection by a variety of parasites. For example, animals on dairy and poultry farms are highly susceptible to enteropathogens, which can cause a high degree of illness and even death, primarily among young animals. To avoid infection, there is a need for efficient sterilization.

Cryptosporidiosis is a diarrheal disease caused by microscopic parasites of the genus Cryptosporidium. Once an animal or person is infected, the parasite lives in the intestine and passes in the stool. The parasite is protected by a chlorine resistant outer coating that allows it to survive outside the body for extended periods.

The compositions of the invention were tested on calves and poultry. In a preliminary experiment, it was found that composition 23, at a final concentration of 5%, is efficient in destroying oocysts of Cryptosporidia under laboratory conditions. Furthermore, it was found that composition 23, at a final concentration of 4%, completely prevents the growth of Salmonella in a test tube. Prior to examining the effectiveness of this composition under field conditions, a safety test was conducted using a 5% aqueous solution of composition 23 on cattle and chickens.

Objective: The purpose of the experiment was to examine the safety of 5% composition of composition 23 on calves and chickens. Methods and materials; Two calves aged two months and 10 chickens, 6 weeks old, were used in the experiment. The bedding upon which the calves and chickens were housed was sprayed three times every two days with a preparation of 5% composition 23. After the final spraying, the animals were clinically examined daily over a period of two weeks. Results: The composition was harmless to the animals. No topical or systemic reaction to the composition was observed.

Example 8: Disinfection of cattle suffering from Cryptospσridiosis 1. Introduction Cryptosporidiosis is very widespread in dairy farms and is caused by a single cell parasite Cryptosporidium parvum. The parasite multiplies in the small intestine and causes illness in calves aged 1-2 weeks. Signs of the disease include a lack of appetite, watery diarrhea and as a result of this, dehydration and sometimes even death, especially when the contagion is also accompanied with other enteropathogens (rota, corona, E. CoIi, salmonella). No effective medical preparations have been found to thwart the multiplication of these parasites in the intestine, which makes the disease very hard to prevent. Furthermore, the sterilization compositions are not sufficiently effective to prevent contagion.

Composition 23 has been found to be effective in destroying the Cryptosporidia under laboratory conditions. Exposure of the parasite to concentrations of 1% and 5%, caused a precipitous drop in the number of oocysts already after a short exposure of half an hour (39% up to 53%), and with an exposure or 6 hours duration at these concentrations, a very significant reduction was observed of 71% up to 88%. From these results, composition 23 is effective in destroying the oocysts of Cryptosporidia in the test tube. A field experiment was conducted to its efficacy at a dairy farm suffering from a Cryptosporidia epidemic.The objective of this experiment was to examine the influence of composition of the present invention on Cryptosporidia infection among calves. Methods and materials: The experiment was conducted in a cowshed containing 300 milk cows. The calves were immediately separated after birth by transferring them to pens in the suckling shed. These pens rest in three rows on the concrete floor and are bedded with straw to a thickness of 5- 10cm. A distance of about 50cm separates the pens from each other. The distance between the rows is about 1-1.5m. The calves were kept in the pens for about two months and thereafter transferred to the fields for grazing. This method allows the natural spread of infection to all the calves, since 8-14 day old calves excrete Cryptosporidia oocysts, which serve as a source of infection for the new calves that are introduced to the suckling shed after birth. To determine the level of natural infection in the farm, fecal samples were examined from ten calves aged 5-15 days for the presence of Cryptosporidia oocysts (control group No.1).

Nine additional calves were tested (control group No.2) prior to the spraying of the bedding with a preparation of composition 23, and 13 other calves (experimental group No.3) after the bedding had been sprayed with a 5% solution of composition 23. The bedding in the pens and their environs, was sprayed daily for a full month, from the introduction of the first calves from the experimental group, until the end of the excretion of oocysts in the feces of all the calves.

The calves were followed-up every day from age 5 to 16 days for the appearance of clinical signs of Cryptosporidiosis and for the presence and quantity of the oocysts in samples of feces taken from each calf separately. Twice during the duration of the experiment, fecal samples from several of the calves in the experiment were examined for the presence of other enteropathogens (bacterial and viral).

Fecal samples were analyzed using the Zihal-Nelson method. In summary, five gram (5g) of feces were dissolved in 20 ml of water, mixed and filtered through a 100- mesh filter. After mixing the solution, 10 microliters (μl) were transferred to a microscope slide. After drying, the plate underwent fixing with methanol for two minutes, stained with concentrated Carbol fuchsin for 30 minutes, removing the staining with a 10% solution OfH₂SO₄ for about 10 seconds and an additional staining with a 0.5% solution of Brilliant green. With this method of staining, Cryptosporidia oocysts were identified under the microscope at a magnification of 400X. The oocysts were seen colored red on a green background on the slide. A calculation of the oocysts per gram of feces was made by multiplying the number of oocysts counted on the complete slide by 500 (the dilution factor). The efficiency of the treatment was determined by comparing the quantity of oocysts excreted by the calves before and after spraying with a 5% solution of composition 23. Results and discussion: The excretion of Cryptosporidia oocysts in calves before and after spraying the bedding with a 5% solution of composition 23. Results for the presence of Cryptosporidia oocysts in feces are detailed in Table 9. Infection with the Cryptosporidia worms was diagnosed in all the calves from the three groups (2 groups before and 1 group after spraying). The calves from the control group (before spraying) began to excrete oocysts at the age of 5-6 days, and excreted on average between 1.25 x 10⁶ - 1.72 x 10⁶ oocysts per gram of feces at the age of 7-8 days and an average of 1.96 x 10⁶ - 4.71 x 10⁶ oocysts per gram of feces at the age 9-12 days. At the age of 13-14 days, a gradual decrease was observed in the excretion of oocysts. At the age of 15 days, most of the calves had ceased to excrete oocysts apart from 2 calves from group No.1, which excrete 10 x 10⁶ and 1.85 x 10⁶ respectively.

Calves from the experimental group, significantly excreted fewer oocysts compared with the calves in the control groups. They began to excrete oocysts at the age of 6-7 days. They excreted only 0.42 x 10⁶ - 0.64 x 10⁶ at the age of 9-11 days (the peak excrete period) and most of the calves ceased to excrete oocysts upon achieving an age of 13 days. The cumulative number of oocysts excrete d by the calves in the experimental group, at all periods of follow-up, was 7.4 times less than the calves in the control group (2.4 x 10⁶ compared with 17.6 x 10⁶ and 17.5 x 10⁶ on average oocysts per gram of feces).

Clinical follow-up: A clinical examination of most of the calves from the control groups showed characteristic signs of Cryptosporidiosis: a lack of appetite, weakness, loss of weight and watery diarrhea especially on the peak days of excreting the oocysts. Calves from the control groups suffered from diarrhea for an average of 7 days. Two calves from the first group died as a result of dehydration at the age of 13 and 14 days. From an examination of feces samples from these calves, no other enteropathogens were observed and one may assume that these calves died from the infection with

Cryptosporidia only. In calves from the experimental group, only light diarrhea was observed which continued for an average of 5 days. Only 2 calves of the 13 in the experimental group showed any signs of watery diarrhea and only for 1-2 days. On the other hand, 15 out of the 19 calves in the control group suffered from watery diarrhea for 3 days on average. From tests for other enteropathogens, a rotavirus infection was diagnosed in 3 out of the 10 calves examined during the experiment. No changes or reactions, either local or general, were observed in any of the calves during the experimental period associated with spraying with a 5% solution of composition 23.

The present experiment results show that the spraying of the bedding of the suckling shed with a 5% solution of composition 23 eradicated a significant proportion of the Cryptosporidia in the cowshed, which is similar to the findings obtained in the lab experiments. The significant reduction in the number of the oocysts owing to spraying composition 23 preparation, resulted in mild infection and during the course of a short illness in the calves from the experimental group, no signs were seen of watery diarrhea, a lack of appetite or dehydration that had been observed in most of the calves in the control groups. Calves from the experimental group excreted 7.4 times fewer oocysts than the control groups, which contributed to a reduction in the level of infection in the suckling shed. Furthermore, it seems that the continued spraying with composition 23, could have contributed to an additional reduction in the level of infection with Cryptosporidia in the cowshed.

Table 9: Average excretion of oocysts in the groups of calves

To conclude, the efficiency of the preparation, as expressed in the experiment described above, is a viable solution for treating calves suffering from a severe outbreak of Cryptosporidiosis. Example 9: Bromide residues from the present composition in soil water runoff and the atmosphere.

Experiments to identify bromine release into the atmosphere and runoff water and bromide residues in the soil following soil disinfection with composition 21 were conducted by an independent laboratory.

Methods: Prior to application of the composition of the present invention, the soil was tested for residual bromide.

Tests were conducted at three different concentrations of the composition, at 750, 1,500 and 3,000 PPM. Three repeat experiments were conducted at all concentrations in order to obtain statistical reliability (a total of 36 tests including a control with 3 repetitions).

The test was conducted within a glove chamber/sealed air chamber into which were placed:

40-cm long, 4-inch diameter columns. Eluate from the column was collected into a collection vessel that was positioned outside of the sealed air chamber. Furthermore, an air spin pump was installed to compress the spun air into an aqueous fixing solution that would absorb the bromine from the air.

Composition 21, at a suitable concentration was blended with the top 10 cm of soil in the column (the lower 30cm of soil in the column was not treated by the composition).

Within the sealed air chamber, the column was wetted with water (eluent), eluate collection was started and the air spin pump was operated for a period of 24 hours.

The eluate was collected and sent for bromine concentration testing.

The air pump within the sealed chamber was operated for 12 hours, at the end of which, the fixing solution was sent to evaluate the bromine concentration.

' The soil from the column was sent for extraction to determine the concentration of bromide.

1. Calculation of quantity of composition for the column:

1.1 For the treatment of 1 ,000 square meters, 200 kg of composition are required (25% active ingredient). 1.2 The internal area of a 4" diameter column is 0.00810 square meters. The volume of the sand in the column is 0.0032 cubic meters or 3.2 liters

The quantity of composition required for the treatment of the (internal surface) volume of the column is 15 g. 1.3 Additional concentrations of composition: 3 g and 4.5 g for the column.

1.4 A quantity of the composition must be mixed with 1,000 cc of soil of the chamra-sand type.

2. Calculation of quantity of water per column:

2.1 1,000 m² of solids in 40,000 liters after the application of composition 2.2 Volume of the column is 3.2 liters.

2.3 Quantity of water required as eluent is about 1.6 liters water.

2.4 Quantity of eluate anticipated: 500 cc.

3. Materials

3.1 Preparation: composition 21 diluted 1 :4 with dry calcium carbonate. 3.2 Soil: Chamra-sand (garden fill sand) or heavy clay soil.

3.3 Water: Haifa Bay area mains tap water.

3.4 Fixing solution: a solution of SMBS (sodium meta-bisulfate) at a concentration of 0.05%.

4. Lab testing methods for bromide concentration: An independent laboratory using an ion chromatograph conducted the tests.

Calculations of volume and quantities are summarized in table 10:

Table 10

Process of testing -Running the control: Three liters of sand were introduced into the column within a plastic sheath.

The column was sealed, and two liters of tap water were run into it using an electric feed pump. After adding the water, the spin taps were opened and the pump was operated for 20 hours. At the end of the spinning, the soil was extracted. All of the samples were sent to an independent laboratory for testing.

Sample 1 : composition concentration equivalent to 200 kg active ingredient/1, 000 m²: 1.5g for the column:

1 gram of active ingredient of composition 21 diluted (dry dilution; cut) 4 fold in calcium carbonate were prepared.

1.5 g of the preparation was mixed with sand to the top 10cm of the column. Water was fed into the column and the experiment proceeded as above (control).

Sample 2: composition concentration of 400 kg active ingredient/ 1,000 m²: 3.0 grams for the column: 3g of the preparation was mixed with sand to the top 10cm of the column. Water was fed into the column and the experiment proceeded as above (control).

Sample 3: composition concentration of 800 kg active ingredient/1, 000 m²: 6.0 grams for the column:

6 g of the preparation was mixed with sand to the top 10cm of the column. Water was fed into the column and the experiment proceeded as above (control).

All of the samples were marked accordingly sent to the independent lab for analysis. Experiments were repeated with columns filled with clay.

Results: Residues of bromide in run-off water, air and soil after applying composition of the invention. Measurements:

Volume of soil in column - 3 liters. Volume of eluent - 2 liters.

Volume of eluate: from sandy soil - 1 liter; from clay soil - 0.25 liter. Volume of fixing solution in the gas trap -0.5 liter. Concentration of SMBS in the sample water - 0.05% Duration of air spin after application- 20 hours.

Method of soil extraction - 20Og of soil (damp) will be mixed with 0.5 liter of SMBS solution and sent for bromide assay. Method of preparing the 0.05% SMBS solution: 2.5g of sodium meta-bi-sulfite powder were added into 5 liter of distilled water and mixed for 10 minutes by an electric magnetic mixer. Quantities of composition 21 for experiment purposes were 1.5g, 3g and 6g, for a column with sand at a volume of 3 liters.

Discussion and conclusions: Bromides in the air: Testing the air for bromides showed that the concentration of bromides in the air is lower than 0.1 PPM. This result indicates that the composition tested does not create volatile bromide compounds. Therefore the composition obviates the danger of damage to the ozone layer, attendant with the use of methyl bromide fumigation.

Bromides in the soil: To tests for residues in the soil show that the level is so low that it is not possible to identify any residues .

Bromides in run-off water: The level of bromides found in the eluate is proportional to the quantity of the composition fed into the column. The quality of the composition used in the experiment was at a concentration of 100% active ingredient. In the field, a composition whose concentration of active ingredient is 20% (diluted about 1 :5), so that the data obtained regarding the concentration of bromides in run-off water are higher by a factor of 5 of the quantities produced owing to the ordinary application of the material. In soil whose depth is greater than 40cm, the level drops so that practically no bromide would reach the underground water reserves

Table 11 shows the bromide concentrations in run-off water, soil and air after sterilizing soil with a composition of the invention. Table 11 : Bromide concentrations

W = eluate, G = soil, A = air, nd= not determined

The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific embodiments without undue experimentation and without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. The means, materials and steps for carrying out various disclosed functions may take a variety of alternative forms without departing from the spirit and scope of the invention.

Claims

1. A dry, water-soluble, non-phytotoxic biocidal composition comprising:

(a) from about 3% wt/wt to about 20% wt/wt of a water-soluble mixture of at least two inorganic halides; (b) at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions;

(c) sulphamic acid;

(d) at least one non-reducing organic acid;

(e) at least one anhydrous alkali metal phosphate; (f) at least one excipient selected from the group consisting of non-organic inert chemical filler, surfactant and a combination thereof; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5. 2. The composition according to claim 1 comprising:

(b) at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions; (c) sulphamic acid;

(d) at least one non-reducing organic acid;

(e) at least one anhydrous alkali metal phosphate;

(f) from about 1% wt/wt to about 61% wt/wt of at least one non- organic inert chemical filler; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.

2 and 5.5.

3. A composition according to claim 1 comprising: (a) from about 3% wt/wt to about 20% wt/wt of a water-soluble mixture of at least two inorganic halides;

(d) at least one non-reducing organic acid;

(e) at least one anhydrous alkali metal phosphate;

(f) from about 5% wt/wt to about 20% wt/wt surfactant; and wherein the pH of a 1% by weight aqueous solution prepared from said

4. The composition according to claim 1 wherein the inorganic halide mixture comprises two or more halides selected from the group consisting of potassium chloride, potassium bromide, potassium iodide, sodium bromide, sodium chloride and sodium iodide.

5. The composition according to claim 4 where the inorganic halide mixture comprises a mixture selected from the group consisting of potassium chloride and sodium bromide; sodium chloride and potassium bromide; sodium chloride and sodium bromide; and potassium chloride and potassium bromide.

6. A composition according to claim 5, wherein the inorganic halide mixture comprises a mixture of potassium chloride and sodium bromide.

7. The composition according to claim 1 wherein the oxidizing agent is selected from the group consisting of a persulfate compound and a peroxyphthalate compound.

8. The composition according to claim 7 wherein the persulfate compound is potassium hydrogen peroxymonosulfate.

9. The composition according to claim 1, wherein the non-reducing organic acid is selected from the group consisting of malic acid, adipic acid, succinic acid and combinations thereof.

10. The composition according to claim 1, wherein the anhydrous alkali metal phosphate is selected from the group consisting of sodium hexametaphosphate, tetrasodium diphosphate, tetrasodium pyrophosphate, mono- sodium phosphate, di- sodium phosphate, tri-sodium phosphate, mono-potassium phosphate (MKP), di- potassium phosphate, tri-potassium phosphate and combinations thereof.

11. The composition according to claim 10 wherein the anhydrous metal phosphate is MKP.

12. The composition according to claim 2 wherein the non-organic inert chemical filler is selected from calcium carbonate and sodium carbonate.

13. The composition according to claim 2 further comprising surfactant.

14. The composition according to any one of claims 3 or 13 wherein the surfactant is selected from the group consisting of lauryl ether sulfates, ethylene oxide, propylene oxide aklyl phenol condensates, polyglycol ethers of fatty alcohols, fatty acid ethylene oxide condensates, polyglycol ethers of alkyl phenols, fatty alcohol ethoxylates and combinations thereof.

15. The composition according to claim 1 comprising

(f) from about 10% wt/wt to about 40% wt/wt non-organic inert chemical filler; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5.

16. A composition according to claim 1 comprising: (a) about 5% wt/wt to about 15% wt/wt of a water-soluble mixture of at least two inorganic halides; (b) from about 35% wt/wt to about 55% wt/wt of at least one oxidizing agent which, in aqueous solution, reacts with the halide to generate hypohalite ions; (c) from about 5% wt/wt to about 8% wt/wt sulphamic acid;

(d) from about 8% wt/wt to about 10% wt/wt of at least one non-reducing organic acid;

(e) from about 10% wt/wt to about 20% wt/wt of at least one anhydrous alkali metal phosphate;

(f) from about 10% wt/wt to about 40% wt/wt of nonorganic inert chemical filler; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5.

17. A composition according to claim 1 comprising:

(a) about 10% wt/wt mixture of potassium chloride and sodium bromide; (b) about 50% wt/wt of potassium hydrogen peroxymonosulfate;

(c) about 5% wt/wt sulphamic acid;

(d) about 8% wt/wt malic acid;

(e) about 10% wt/wt MKP; (f) about 17% wt/wt of non-organic inert chemical filler; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5.

18. A solid composition for soil disinfestation comprising about 15% wt/wt to about 50% wt/wt of a composition according to any one of claims 2, 12 or 15- 17; and about 50% wt/wt to about 85% wt/wt of an inorganic, inert chemical filler.

19. The composition according to claim 2 comprising

(f) from about 5% wt/wt to about 20% surfactant; and wherein the pH of a 1% by weight aqueous solution prepared from said composition being between 1.2 and 5.5.

20. An aqueous solution comprising about 0.05% to about 20% wt/v of a composition according to any one of claims 1-3 or 15-17.

21. A method for soil disinfestation comprising the step of: a) applying to the soil a biocidal-effective amount of the composition according to any one of claims 2, 12, 15-17, or 18.

22. The method according to claim 21, further comprising the step of: b) blending said composition into the soil.

23. A method for seed disinfection comprising the steps of: a) preparing a dilute aqueous solution of the composition according to any one of claims 1-3, 13, 15-17 or 19; and b) wetting the seeds in said solution.

24. The method according to claim 23 wherein the aqueous solution comprises about 0.05% wt/v to about 20% wt/v of said composition.

25. The method according to claim 23 wherein the aqueous solution comprises about 0.1% wt/v to about 2% wt/v of said composition.

26. A method for fish disinfection comprising the steps of: a) preparing a dilute aqueous solution of the composition according to any one of claims 1-3, 13, 15- 17 or 19; and , b) exposing fish to said solution.