Classifications

• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
  • A01N37/16—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing the group; Thio analogues thereof
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01M—CATCHING, TRAPPING OR SCARING OF ANIMALS; APPARATUS FOR THE DESTRUCTION OF NOXIOUS ANIMALS OR NOXIOUS PLANTS
  • A01M17/00—Apparatus for the destruction of vermin in soil or in foodstuffs
  • A01M17/002—Injection of toxic gases or fluids into the soil
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
  • A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
  • A61L2/18—Liquid substances or solutions comprising solids or dissolved gases
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
  • A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
  • A61L2/18—Liquid substances or solutions comprising solids or dissolved gases
  • A61L2/186—Peroxide solutions
• • A61L2103/05—

Definitions

• the present invention relates to compositions and methods for soil disinfection.
• the compositions and methods are useful against a variety of plant pathogens, including nematodes, bacteria, and fungi.
• methyl bromide (CH 3 Br) had been the most widely used and most universal fumigant in the world. It is known for being extremely effective as a nematicide, insecticide, fungicide, and herbicide. Methyl bromide has been used extensively as a soil fumigant, a commodity quarantine treatment for exports and imports, as a pesticide on numerous crops, and as a structural fumigant applied to building surfaces. However, methyl bromide has contributed to the depletion of the ozone layer in the stratosphere. In accord with the Montreal Protocol, the import and manufacture of methyl bromide in the United States and other developed countries was banned in 2005. For developing countries, the reductions were more gradual and the phase-out was delayed until 2015.
• Organic peracids such as peracetic acid (PAA) are potent biocides. They have had limited utility as soil sterilants, in part, because the relatively high concentration of organic matter in soil may inactivate the peracids.
• PAA peracetic acid
• U.S. Pat. No. 5,168,655 discloses an aqueous solution comprising peracetic acid being made in-situ by mixing acetic acid, hydrogen peroxide, 2,6-pyridine dicarboxylic acid, dodecyl benzene sulphonic acid, and water.
• the solution is used in an irrigation system to partially control bacteria, fungal, spores, yeast, and molds in hydroponic substrates.
• U.S. Pat. No. 5,168,655 requires that the substrate contain only a minimal amount of organic matter because excessive amounts of organic matter, such as those in soil, are believed to inactivate the peracetic acid.
• Aromatic alkyl sulphonic acid specifically odecyl benzene sulphonic acid, is disclosed as a wetting agent in the solution in U.S. Pat. No. 5,168,655.
• EP 0035800 discloses an aqueous solution containing hydrogen peroxide and/or peracids having 1 to 4 carbon as soil treatment agent to control phytopathogenic harmful organisms, such as fungi, bacteria, and nematodes.
• the present invention relates to and aqueous composition
• a peracid can be peracetic acid.
• the peracetic acid can be an aqueous equilibrium solution comprising a weight ratio of peracetic acid:hydrogen peroxide between 1:0.01 to 1:14 and a weight ratio of peracetic acid:acetic acid between 1:0.2 to 1:19.
• the peracetic acid concentration can be from about 3000 ppm to about 25,000 ppm.
• the polyoxyethylene alkylether carboxylic acid concentration can be from about 250 ppm to about 25,000 ppm.
• the peracid can be peracetic acid.
• the pathogenic microorganism can be a nematode, bacteria, or fungus.
• the agricultural medium can include soil, sand, or a synthetic growth medium.
• the agricultural medium can be contacted with the composition by spraying, drenching, injecting, sprinkling, or infusing the composition into the agricultural medium.
• the aqueous solution is applied at about 1.5 mL/cm 2 of agricultural medium.
• machine When only a single machine is illustrated, the term “machine” shall also be taken to include any collection of machines that individually or jointly execute a set (or multiple sets) of instructions to perform any one or more of the methodologies discussed herein.
• means-plus-function clauses if used, are intended to cover the structures described, suggested, or rendered obvious by the written description or drawings for performing the recited function, including not only structural equivalents but also equivalent structures.
• the present invention is directed to formulations for treatment of soil to control plant parasitic nematodes and soil-borne plant pathogens, for example, bacteria and fungi.
• the formulations include (i) peracids, such as peracetic acid and (ii) anionic surfactants, such as capryleth-9 carboxylic acid.
• Peracetic acid is typically employed in the form of an aqueous equilibrium mixture of acetic acid (AA), hydrogen peroxide (H 2 O 2 ) and peracetic acid (PAA).
• AA acetic acid
• H 2 O 2 hydrogen peroxide
• PAA peracetic acid
• the weight ratios of these components may vary greatly, depending upon the particular grade of PAA employed.
• grades of PAA which may be employed are those having a weight ratio of PAA:hydrogen peroxide between 1:0.01 to 1:14 and a weight ratio of PAA:acetic acid between 1:0.2 to 1:19.
• peracetic acid solutions include 5% PAA with 22% H 2 O 2 and 10.5% AA, 15% PAA with 10% H 2 O 2 and 35% AA, 15% PAA with 23% H 2 O 2 and 16% AA, 22% PAA with 10% H 2 O 2 and 35% AA, and 35% PAA with 6.5% H 2 O 2 and 40% AA.
• the anionic surfactant can be a polyoxyethylene alkylether carboxylic acid, or a related anionic surfactant, including, for example, polyoxyethylene octyl ether carboxylic acid, polyoxyethylene(8) octyl ether carboxylic acid, polyoxyethylene(10) oleyl ether carboxylic acid, polyoxyethylene(10) lauryl ether carboxylic acid, polyoxyethylene(3) lauryl ether carboxylic acid, polyoxyethylene(5) lauryl ether carboxylic acid, polyoxyethylene(7) lauryl ether carboxylic acid, polyoxyethylene(2) oleyl ether carboxylic acid, polyoxyethylene(5) oleyl ether carboxylic acid, polyoxyethylene(9) oleyl ether carboxylic acid, and salts of above said polyoxyethylene alkylether carboxylic acids in sodium, potassium, or other cation forms.
• the anionic surfactant is polyoxyethylene alkylether carboxylic acid, which is also known as Capryleth-9 Carboxy
• compositions of the invention can include a peracid, for example peracetic acid, and an anionic surfactant, for example, capryleth-9-carboxylic acid (C9CA).
• a peracid for example peracetic acid
• an anionic surfactant for example, capryleth-9-carboxylic acid (C9CA).
• the peracid and the anionic surfactant can be diluted with water to the desired concentrations and combined at the point of use.
• the peracid and the anionic surfactants can be combined to form a mixture and the mixture can be diluted with water before use.
• the peracid and the anionic surfactant can be combined and stored before use or they can be combined and used directly.
• compositions employed in the formulation of the invention can further include or exclude sequestriants such as dipicolinic acid and 1-hydroxyethylidene-1,1,-diphosphonic acid, as well as other ingredients such as mineral acid catalysts (sulfuric, nitric, or phosphoric acids); and short chain fatty esters (C6-C12) forming mixed peracids in solution.
• sequestriants such as dipicolinic acid and 1-hydroxyethylidene-1,1,-diphosphonic acid
• other ingredients such as mineral acid catalysts (sulfuric, nitric, or phosphoric acids); and short chain fatty esters (C6-C12) forming mixed peracids in solution.
• compositions employed in the formulation of the invention may further include or exclude one or more additional oxidants selected from the group consisting of chloroperbenzoic acid, perheptanoic acid, peroctanoic acid, perdecanoic acid, performic acid, percitric acid, perglycolic acid, perlactic acid and perbenzoic acid.
• additional oxidants selected from the group consisting of chloroperbenzoic acid, perheptanoic acid, peroctanoic acid, perdecanoic acid, performic acid, percitric acid, perglycolic acid, perlactic acid and perbenzoic acid.
• compositions are diluted into water before use, and the diluted formulation can be applied using application techniques and equipment for soil fumigants in liquid form, such as trench applications, handgun applications, shank (chisel) applications, sweep or blade applications, drench application, and chemigation.
• application techniques and equipment for soil fumigants in liquid form such as trench applications, handgun applications, shank (chisel) applications, sweep or blade applications, drench application, and chemigation.
• the present formulations may be used to control plant parasitic nematodes and soil-borne plant pathogens of bacteria and fungi.
• Exemplary plant parasitic nematodes include root knot nematodes such as Meloidogyne hapla, Meloidogyne incognita, Meloidogyne enterolobii and Meloidogyne mayaguensis ; cyst nematodes such as soybean cyst nematodes ( Heterodera glycines ); potato cyst nematodes ( Globodera pallida and G. rostochiensis ) and cereal cyst nematodes ( Heterodera avenae and H.
• root lesion nematodes such as ( Pratylenchus spp., including P. penetrans, P. thornei, P. neglectus, P. zeae, P. vulnus and P. coffeae ; and the burrowing nematode, Radopholus similis.
• Exemplary soil bacteria include Bacillus species, for example Bacillus mycoides .
• Exemplary soil fungi include Aspergillus species, for example Aspergillus niger.
• Peracetic acid formulations were evaluated for nematocidal and bactericidal activity.
• Formaldehyde was used as a positive control.
• the test conditions are described below and the results are summarized in Tables 1, 2, and 3.
• Test method A peracetic acid stock solution containing 5% PAA and 22% H 2 O 2 (VigorOx® from PeroxyChem, LLC) was used for methods that specified 1750 ppm PAA. A peracetic acid stock solution containing 15% PAA and 10% H 2 O 2 (VigorOx SP-15) was used for methods that specified 3525 ppm PAA. A stock solution of formaldehyde (37% in 10-15% methanol) was used to prepare a 1% aqueous solution of formaldehyde (with about 0.3% methanol).
• the Test Medium was: pasteurized 50:50 Loamy sand or pasteurized sand.
• the efficacy of PAA was evaluated in three different Greenhouse Test Assays: 1) the Tomato Root-knot Galling Index Assay using the nematode species Meloidogyne incognita and Meloidogyne hapla; 2) the Root-knot Nematode Egg Hatch Assay using the nematode species Meloidogyne incognita and Meloidogyne hapla; and 3) assay of levels of soil bacteria using Bacillus mycoides.
• the Root-knot Nematode Egg Hatch Assay was carried out essentially as follows. Eggs were collected from roots of infected tomato plants. To release the eggs, the egg masses were placed in a solution of sodium hypochlorite and manually shaken. The solution was centrifuged and the eggs washed with distilled water. Eggs were kept in the dark until needed for the experiments and used in the same day of collection. The experiments were performed in pots filled with pasteurized soil inoculated with about several thousands of individual eggs and then drenched with each treatment solution and controls. The treated soils were covered and incubated for the time periods indicated below. Following the incubation period, the soils were transferred to collect hatched J2 (second stage juveniles). The collected material was observed under the microscope and the number of J2 counted.
• Root Knot Nematode Egg Hatch Assay The results of the Root Knot Nematode Egg Hatch Assay are shown in Table 2. Seedlings were inoculated and cultured in pasteurized Pemberton soil for 14 days. Seedlings were then treated with either formaldehyde, 1750 ppm PAA or 3525 ppm PAA for 13 days. Nematode eggs were extracted from infected roots and their ability to hatch was evaluated. As shown in Table 2, treatment of soil with 3525 ppm PAA resulted in a 51% reduction of the J2 (second stage juvenile) population relativity to control. Treatment of soil with 1750 ppm PAA did not reduce the J2 population relative to untreated controls. Treatment with 1% formaldehyde resulted in a 78% reduction relative to untreated controls.
• the formulations were also assayed for the ability to reduce soil bacteria as shown in Table 3.
• Soil was inoculated with Bacillus mycoides , incubated for five days and then treated with formaldehyde or 1750 PPA for four days. Bacteria were collected from soil and their levels were evaluated based on colony forming units. As shown in Table 3, treatment of soil with 1750 ppm PAA resulted in a 47.5% reaction of colony forming units (CFU's) relative to control. Treatment of soil with 1% formaldehyde resulted in a 42.5% reduction of colony forming units (CFU's) relative to control.
• Peracetic acid formulations were evaluated for nematocidal activity either alone or in combination with C9CA.
• Formaldehyde was used as a positive control.
• Test method A peracetic acid stock solution containing 15% PAA and 10% H 2 O 2 (VigorOx SP-15) was used for methods that specified 3750, 5000, 7500, 10000 ppm PAA.
• Capryleth-9 carboxylic acid (C9CA) was used at concentrations of 5000 and 10000 ppm.
• a stock solution of formaldehyde (37% in 10-15% methanol) was used to prepare a 1% aqueous solution of formaldehyde. The final methanol concentration was about 0.3%.
• the Test Medium was pasteurized Pemberton loamy soil, sand or pasteurized sand.
• the efficacy of PAA was evaluated with the Root-Knot Nematode Egg Hatch Assay using the nematode species Meloidogyne incognita and Meloidogyne hapla.
• Root Knot Nematode Egg Hatch Assay The results of the Root Knot Nematode Egg Hatch Assay are shown in Table 4. Seedlings were inoculated and cultured for 22 days and then treated for 21 days with either PAA alone at various concentrations, C9CA alone at various concentrations, or a combination of PAA and C9CA. Nematode eggs were extracted from infected roots and their ability to hatch was evaluated.
• Example 2 PAA alone at a dosage between 3500 to 10000 ppm provided a moderate reduction in J2 nematode population in soil, which is consistent with the result in Example 1.
• C9CA provided minimal nematicidal activity when used alone at either 5000 ppm or 10000 ppm.
• PAA alone at 5000 ppm produced a 20% reduction in egg hatch.
• C9CA alone did not reduce the nematode population.
• the combination of PAA at 5000 ppm and C9CA at 5000 ppm resulted in a 40% reduction in the nematode population, as measured by Egg Hatch levels.
• Peracetic acid formulations were evaluated for nematocidal activity either alone or in combination with C9CA. Distilled water was used as a control in order to calculate the percentage of egg hatch that occurred in untreated soil.
• Test method A peracetic acid stock solution containing 15% PAA and 10% H 2 O 2 (VigorOx SP-15) was used for methods that specified 3500 and 20,000 ppm PAA.
• the viability of the root-knot nematodes eggs used for each set of experiments in this example was also evaluated. Eggs were incubated with the different solutions in glass dishes in the dark at 25° C. They were observed daily under the microscope to confirm that the eggs were viable and able to hatch in water after 18 hr/24 hr of incubation. These observations confirmed that the eggs used for each set of soil experiments were viable.
• Root Knot Nematode Egg Hatch Assay The results of the Root Knot Nematode Egg Hatch Assay are shown in Table 5. Seedlings were inoculated and cultured for 22 days and then treated for 21 days with either PAA alone at various concentrations, C9CA alone at various concentrations, or a combination of PAA and C9CA. Nematode eggs were extracted from infected roots and their ability to hatch was evaluated.
• soil treatment with PAA alone at 20,000 ppm had an inhibitory effect on egg hatching.
• Soil treatment with PAA alone at 20,000 ppm reduced egg hatching to 53% of distilled water treated control levels.
• Soil treatment with C9CA alone also had a modest inhibitory effect on egg hatching.
• Soil treatment with C9CA alone at 250 ppm reduced egg hatching to 66% of distilled water treated control levels. Consistent with the results shown in Example 2, when PAA and C9CA were used together, the effect on reduction of nematode population was substantially greater than the effect of either agent alone. Moreover, the inhibitory effect was dose-dependent.
• Soil treatment with 3500 ppm PAA+250 ppm C9CA reduced egg hatching to 45% of distilled water treated control levels.
• Soil treatment at higher doses, (20000 ppm PAA+10000 ppm C9CA) reduced the percentage of egg hatching to only 11% of distilled water treated controls. This synergistic effect was unexpected.
• Peracetic acid formulations were evaluated for sporicidal activity.
• Test method A peracetic acid stock solution containing 15% PAA and 10% H 2 O 2 (VigorOx SP-15) was used for methods that specified 3525 ppm PAA. Deionized water was used as a control.
• the Test Medium was pasteurized Pemberton soil.
• the spore inoculum was Bacillus subtilis ATCC 19659. Spores were counted as follows: Soil was pasteurized to remove background microbial content. The soil was distributed into test cells. Concentrated Bacillus subtilis ATCC 19659 spore suspension was diluted in Butterfield's buffer and used as the inoculum in this test. For titer determination, the inoculum was diluted in Butterfield's buffer and plated on Petrifilm APC. In order to test for antimicrobial efficacy following treatments, soil was scooped from the cell and measured into a sterile plastic centrifuge tube.
• Deionized water was then added to each soil sample.
• the tubes were then capped, shaken to suspend soil, and vortexed at maximum speed using the lab vortex. The samples were allowed to settle for several seconds, and then 2 mL of fluid was removed from the tube. One ml of the removed sample was diluted in 9 mL Butterfield's buffer. The remainder of the removed sample was plated onto Petrifilm APC. The diluted sample was further serially diluted and plated on APC. The plates were all incubated at 30 ⁇ 2° C. for 2-5 days and then colonies on the plates were counted.
• PAA was evaluated either alone or in combination with C9CA for fungicidal activity.
• Test method A peracetic acid stock solution containing 15% PAA and 10% H 2 O 2 (VigorOx SP-15) was used for methods that specified 3500 and 20,000 ppm PAA.
• the Test Medium was pasteurized Pemberton soil.
• the spore inoculum was Aspergillus niger ATCC 16404.
• the fungicidal activity of the compositions was evaluated as follows: Soil was pasteurized to remove background microbial content. The soil was the filled into test cells. Aspergillus niger ATCC 16404 was grown on Potato Dextrose Agar (PDA) plates for 5 days at 35° C.
• PDA Potato Dextrose Agar
• a culture for test was prepared by washing a plate with Butterfield's buffer, grinding in a sterile tissue grinder, and filtering through a cell filter to remove hyphae and debris.
• the suspension was diluted 1:100 in Butterfield's buffer and was added to the soil as the inoculum.
• the inoculum was enumerated on Petrifilm APC to determine the titer.
• soil was scooped from the cell and weighed into a sterile plastic cup.
• Sterile deionized water was added to the cup using a serological pipette.
• the cups were then capped, shaken to suspend soil, and vortexed at maximum speed using the lab vortex.
• the samples were allowed to settle for several seconds, and then 2 mL of fluid was removed from the tube.
• One ml of the removed fluid was diluted in 9 mL Butterfield's buffer.
• the remaining removed fluid was plated onto Petrifilm Petrifilm YM.
• the diluted sample was further serially diluted and plated to 10 ⁇ 3 on YM.
• the plates were all incubated at 35 ⁇ 2° C. for 3 to 4 days and colonies were counted.
• PAA alone provided a Log 10 reduction of more than 2 at even at the short treatment time of one hour. Aspergillus niger was completely eliminated after 24 hrs of treatment. Doubling the PAA dosage to 7000 ppm resulted in complete elimination even after only one hour of treatment.

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