METHOD FOR CONTROLLING PATHOGENS AND/OR PREVENTING DISEASES RESULTING FROM THE PRESENCE OF THE PATHOGENS IN AND/OR ON SEEDS

Abstract

A method for the control of pathogens and/or the prevention of diseases associated with the presence of said pathogens in and/or on seeds, said method comprising the steps of contacting the seeds with a sanitizing composition comprising at least one agriculturally acceptable sanitizing agent, water; and optionally at least one agriculturally acceptable alcohol; heating/drying the seeds at a temperature varying from 160 F. to 230 F. for a period of time varying from 9 to 16 minutes for a period of time that prevents the core of the seeds heated to reach 70 C.; and optionally a complementary drying of the seeds at a temperature <70 C.

Claims

1. A method for the treatment of seeds, said treatment allowing to control the amount of pathogens in and/or on the seeds and/or allowing to prevent diseases associated with the presence of said pathogens in and/or on seeds and/or parts of seeds, the method comprising: contacting the seeds with a sanitizing composition comprising: at least one oxidizer; water; and at least one alcohol, to produce pre-treated seeds; and drying the pre-treated seeds in a drying environment having a first temperature of between 160 F. and 230 F. while preventing a core of the seeds from reaching a temperature 158 F.

2. The method of claim 1, wherein the seeds have an initial moisture content and contacting the seeds with the sanitizing composition results in the pre-treated seeds having a moisture content greater than the initial moisture content.

3. The method of claim 1, wherein the drying of the pre-treated seeds is performed in a continuous dyer and comprises regulating evaporation to prevent cooking of the core of the seeds.

4. The method of claim 1, wherein the drying is performed for a period of between 9 and 16 minutes, or about 2 minutes.

5. The method of claim 1, further comprising at least one subsequent drying step wherein the drying environment has a subsequent temperature that is lower than the first temperature.

6. The method of claim 5, wherein the subsequent temperature is <158 F.

7. The method of claim 5, wherein the at least one subsequent drying step comprises a second drying step performed at a second temperature that is <158 F., followed by a third drying step that is performed at a third temperature that is <120 F.

8. The method of claim 5, wherein the seeds are dried until the seeds reach a moisture content that is close to an initial moisture content of the seeds, the initial moisture content being less than 10 wt. %.

9. The method of claim 1, wherein the oxidizer is a liquid peracetic acid, in-situ generated peracetic acid from powder precursors, liquid hydrogen peroxide, hydrogen peroxide released from a powder persalt, or mixtures thereof.

10. The method of claim 1, wherein the alcohol is a glycol ether, a propylene glycol, an ethylene glycol, a C.sub.1-C.sub.6 linear alkanol or a C.sub.3-C.sub.6 branched alkanol.

11. The method of claim 1, wherein the seeds are provided with their natural envelope, shell or hard shell.

12. The method of claim 1, wherein the sanitizing composition comprises from 1 wt. % to 10 wt. % of the at least one oxidizer, from 1 wt. % to 40 wt. % of the at least one alcohol, and from 50 wt. % to 98 wt. % of water.

13. The method of claim 1, wherein the drying of the pre-treated seeds prevents rancidity or changes in the sensory attributes of the seeds.

14. The method of claim 1, wherein the pathogens are viruses, bacteria, fungi, yeasts or moulds, and wherein: the bacteria are selected from the group consisting of E. Coli, Listeria monocytogenes, Salmonella spp. and E. faecium, Agrobacterium spp., Burkholderia spp., Clavibacter spp., Corynebacterium spp., Erwinia spp., Pseudomonas spp., Ralstonia spp., Rhizomonas spp., Xanthomonas spp., and Xylella spp.; and the fungi are selected from the group consisting of Albugo spp., Alternaria spp., Armillaria spp., Aspergillus spp., Athelia spp., Bipolaris spp., Botryosphaeria spp., Botryotinia spp., Botrytis spp., Bremia spp., Capnodium spp., Ceratobasidium spp., Ceratocystis spp., Cercospora spp., Choanephora spp., Claviceps spp., Corynespora spp., Cronartium spp., Cryphonectria spp., Cylindrocladium spp., Cytospora spp., Diaporthe spp., Diplodia spp., Dreschlera spp., Elsinoe spp., Erexohilum spp., Erysiphe spp., Eutypha spp., Exobasidium spp., Fusarium spp., Gaeumannomyces spp., Gliocladium spp., Gymnosporangium spp., Heterobasidium spp., Hypoxylon spp., Kutilakesa spp., Lophiodermium spp., Magnaporthe spp., Melampsora spp., Monilinia spp., Mycosphaerella spp., Myrothecia spp., Nectriella spp., Nematospora spp., Oidium spp., Olpidium spp., Ophiostoma spp., Penicillium spp., Peronospora spp., Phakospora spp., Phoma spp., Phomopsis spp., Phragmidium spp., Phyllactinia spp., Physoderma spp., Phytophthora spp., Plasmodiophora spp., Plasmopara spp., Pseudoperonospora spp., Puccinia spp., Pythium spp., Rhizoctonia spp., Rhizopus spp., Rhytisma spp., Sclerotinia spp., Sclerotium spp., Spongospora spp., Synchytrium spp., Taphrina spp., Thanatephorus spp., Thielaviopsis spp., Tilletia spp., Uncinula spp., Urocystis spp., Ustilago spp., Valsa spp., Venturia spp., Verticillium spp., and Xylaria spp.

15. The method of claim 1, wherein the pathogen is E. faecium NRL B02354.

16. The method of claim 1, wherein the seeds are: selected from the group consisting of cereals, pseudocereals, nuts, nut-like gymnosperm seeds, cempedak seeds, coffee seeds, egusi seeds, euryale ferox (fox nut) seeds, fluted pumpkin seeds, hemp seed seeds, jackfruit seeds, lotus seeds, Malabar gourd seeds, pumpkin seed seeds, sunflower seed seeds, sesame seeds or Tahini seeds, beans, seeds for sprouting, seed spices, seeds of crops transplantable from greenhouse to field, and seeds of marijuana; or selected from the group consisting of filberts, almond, chia, cashews, walnut, hazelnuts and sunflower kernels.

17. A method for controlling or preventing diseases associated with the presence of pathogens in and/or on seeds or parts thereof, the method comprising: contacting the seeds with a sanitizing composition comprising: at least one oxidizer; water; and at least one alcohol, to produce pre-treated seeds; and drying the pre-treated seeds in a drying environment having a temperature of between 160 F. and 230 F. while preventing a core of the seeds from reaching a temperature 158 F.

18. The method of claim 17, wherein the alcohol is a glycol ether, a propylene glycol, an ethylene glycol, a C.sub.1-C.sub.6 linear alkanol or a C.sub.3-C.sub.6 branched alkanol.

19. The method of claim 17, wherein the sanitizing composition comprises from 1 wt. % to 10 wt. % of the at least one oxidizer, from 1 wt. % to 40 wt. % of the at least one alcohol, and from 50 wt. % to 98 wt. % of water.

20. A method for controlling or preventing diseases associated with the presence of pathogens in and/or on seeds or parts thereof, the method comprising: contacting the seeds with a sanitizing composition comprising at least one oxidizer and water, to produce pre-treated seeds; and drying the pre-treated seeds in a continuous dryer having a temperature of between 160 F. and 230 F. for about 2 minutes, while preventing a core of the seeds from reaching a temperature 158 F., to prevent rancidity or changes in the sensory attributes of the seeds.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0112] The present invention will be better understood with reference to the following drawings:

[0113] FIG. 1 represents the effect on reduction of E. faecium in filberts according to example 1, when treated with either a sanitizing composition containing Neo-Pure sanitizing agent or water at 50 L/t, or no treatment and after drying at three different temperatures (71.1 C., 82.2 C. and 93.3 C.).

EXAMPLES

[0114] The following examples illustrate surprising improvements according to the present invention.

[0115] In the following examples 1 to 3, when a heating/drying step is involved, it was advantageously carried out with a fluid bed dryer of the type known as a Sherwood Scientific Model 501 Fluid Bed Dryer, allowing air to flow through a bed of particles (in the present examples seeds) at controlled temperature and flow rate. The bed of particles was then assumed to be in a fluid-like state (resembling a boiling liquid), and the heating of the air flow (entering the bed of particles) and the managing of the rate of the air flow through the particles, provide a thorough mixing and maximum contact of the particles (e.g. seeds) with the moving air flow while allowing to obtain a heating/drying of particles much quicker drying than with conventional methods.

Example 1

Preparation of a Sanitizing Composition SC1

[0116] The sanitizing composition SC1 comprised a sanitizing agent prepared from a NEO-PURE liquid formulation comprising by weight:

TABLE-US-00001 1) Distilled Water= 34.5% 2) Acetic Acid 92%= 13.5% 3) Sulphuric Acid 96%= 1.1% 4) Hydrogen Peroxide 50%= 50% 5) Dequest 2010 (diphosphonic acid) 0.9% TOTAL: 100%.

[0117] This sanitizing agent generates peracetic acid in an amount of about 5% by weight of peracetic acid with respect to the total weight of the sanitizing agent. The sanitizing composition SC1 is obtained by mixing 10 percent by volume of the above-mentioned NEO-PURE sanitizing agent with 90 percent by volume of water to give the following sanitizing composition. As an example, for providing 50 litres of the sanitizing composition SC1, 5 litres of NEO-PURE are mixed with 45 litres of water. This sanitizing composition is particularly adapted for spraying on seeds.

Effect Between the Sanitizing Composition SC1 and a Subsequent Heating/Drying Step for the Reduction of the Salmonella Surrogate E. faecium NRRL B-2354 in Filberts.

[0118] The objective of this example was to determine the effect of a heating/drying step following a step of spraying of the sanitizing composition SC1 on filberts (shelled hazelnuts).

[0119] Filberts sprayed with the sanitizing composition SC1 were then subjected to a heating/drying step at different temperatures. The heating/drying step was carried out in a Sherwood Scientific Model 501 Fluid Bed Dryer.

Method:

[0120] Several kg of filberts (shelled hazelnuts) were inoculated with a 2% inoculum of E. faecium and mixed thoroughly for 1 min. Then, the inoculated filberts were heated/dried into a fluid bed dryer (Sherwood Scientific Model 501 Fluid Bed Dryer) at 40 C. for 10 min, until the filberts reached their original % moisture content (about 3.3% wt.-%). After, the following treatments were applied (i.e. sprayed) to 1 kg (X3) of the inoculated filberts: [0121] 1) SC1 (at a rate of 50 L/tonne). [0122] 2) Water (at a rate of 50 L/tonne). [0123] 3) No treatment.

[0124] After, the filberts were heated/dried at either: [0125] 1) 71.1 C. (9 min) [0126] 2) 82.2 C. (12 min) [0127] 3) 93.3 C. (16 min)

[0128] The heating/drying time was the time needed for the samples to reach their original % moisture content at each temperature.

[0129] For the E. faecium enumeration, 5 samples of 45 g were taken from the untreated controls (UTCs), the treated samples, and the heated/dried samples.

[0130] Enumeration of E. faecium was done following the procedure described in the FDA Bacteriological Analytical Manual (BAM) (Andrews and Hammack, 2003). The samples were then diluted with buffered peptone water (BPW) (1:2 w/v) in sterile stomacher bags.

[0131] Samples were mixed through shaking vigorously 50 times in a 30 cm (1 ft.) arc with hand. Then, the samples were left stand for 3-5 minutes and shaken vigorously 5 times in a 30 cm arc, just before making serial dilutions.

[0132] Subsequently, 10-fold serial dilutions in a buffered peptone water (BPW) were prepared, and aliquots were plated on Enterococci selective agar (Slanetz & Bartley), followed by incubation at 35 C. for 48 h. Results are reported in log CFU/g.

[0133] The detection limit was 2 CFU/g (0.3 log CFU/g). The average log reduction was determined by subtracting each of the remaining counts of E. faecium after the treatments to the average UTC CFU/g log, and obtaining the average of the 5 values.

Results:

TABLE-US-00002 TABLE 1 Comparison of the effect of the sanitizing composition SC1 (SC1 in the following table) and heat/dry the treatment of filberts alone and combined Product: Filberts Objective: To compare the effect of SC1 and heating/drying on the treatment of filberts alone and combined Avg. Avg. SAMPLE UTC Treated only Log Treated + Dried Log UTC's Drying (rate, Log S Log S red. Std Log Std red. Std % UTC's time drying T) CFU/g dev CFU/g dev CFU/g Dev CFU/g dev CFU/g Dev M.C. a.sub.w (min) SC1, 5.81 0.12 3.25 0.35 2.56 0.35 1.42 0.63 4.39 0.67 3.30 0.36 16 50 L/t, 71.1 C. SC1, 1.60 0.67 4.21 0.67 12 50 L/t, 82.2 C. SC1, 0.78 0.61 5.03 0.61 9 50 L/t, 93.3 C. Water, 5.68 0.35 0.13 0.35 4.24 0.33 1.57 0.33 16 50 L/t, 71.1 C. Water, 4.37 0.06 1.44 0.06 12 50 L/t, 82.2 C. Water, 4.22 0.10 1.59 0.10 9 50 L/t, 93.3 C. No NA NA NA NA 5.65 0.19 0.16 0.19 16 treatment, 71.1 C. No 5.53 0.33 0.28 0.33 12 treatment, 82.2 C. No 5.65 0.10 0.16 0.10 9 treatment, 93.3 C.

TABLE-US-00003 TABLE 2 Complement of information concerning Table 1 regarding the Log reduction (CFU/g) Log reduction (LogCFU/g) Temperature ( C.) 3.9 71.1 82.2 93.3 No treatment 0 0.16 0.28 0.16 Water (50 L/t) 0.13 1.57 1.44 1.59 SC1 (50 L/t) 2.56 4.39 4.21 5.03 SC1 (50 L/t) + Temperature, 2.56 2.72 2.84 2.72 Theoretical

TABLE-US-00004 TABLE 3 Complement of information concerning Table 1 regarding the StDev StDev Temperature ( C.) 3.9 71.1 C. 82.2 C. 93.3 C. No treatment 0 0.19 0.33 0.10 Water 0.35 0.33 0.06 0.10 SC1 0.35 0.67 0.67 0.5

TABLE-US-00005 TABLE 4 Complement of information concerning Table 1 concerning the E. faecium Log reduction (LogCFU/g) Filberts E. faecium Log reduction (LogCFU/g) Temperature ( C.) 3.9 71.1 82.2 93.3 No treatment 0 0 0.16 0.19 0.28 0.33 0.16 0.10 Water (50 L/t) 0.13 0.35 1.57 0.33 1.44 0.06 1.59 0.10 SC-Ex. 1 (50 L/t) 2.56 0.35 .sup.4.39 0.0.67 4.21 0.67 5.58 0.5 SC-Ex. 1 (50 L/t) + 2.56 2.72 2.84 2.72 Temperature (theoretical)

[0134] The graph of FIG. 1 shows the effect on reduction of E. faecium in filberts when treated with either sanitizing composition SC1 or water at a rate of 50 L/t, or no treatment and after drying at three different temperatures (71.1 C., 82.2 C. and 93.3 C.).

[0135] When the inoculated filberts were treated with sanitizing composition SC1, and heated/dried at either of the 3 different temperatures, more than 4 log CFU/g reduction on E. faecium was achieved.

[0136] However, when the filberts were treated with water, the log reductions achieved were below 2 log CFU/g, even at the highest temperature, 93.3 C.

[0137] Furthermore, when the filberts were not treated with any liquid, but heated/dried at the three temperatures, minimal effect in the counts of E. faecium was observed, below 0.3 log CFU/g reduction.

[0138] In addition, when the theoretical value of the log reduction of the sanitizing composition SC1 and temperature alone (SC1 (50 L/t)+temperature (theoretical)) was calculated, the results were at least 1.5 log CFU/g below the values achieved when the sanitizing composition SC1 and temperature were actually combined in the treatment of filberts.

CONCLUSION

[0139] It can be concluded that there is a synergistic effect between the sanitizing composition SC1 and the heating/drying treatment applied during the heating/drying process at different temperatures (71.1 C., 82.2 C. and 93.3 C.) on reduction of E. faecium in filberts.

Example 2

Sanitizing Composition 2 (SC2)

[0140] This sanitizing composition SC2 comprised a sanitizing agent prepared from a NEO-PURE liquid formulation comprising by weight:

TABLE-US-00006 1) Distilled Water= 34.5% 2) Acetic Acid 92%= 13.5% 3) Sulphuric Acid 96%= 1.1% 4) Hydrogen Peroxide 50%= 50% 5) Dequest 2010 (diphosphonic acid)= 0.9% TOTAL: 100%.

[0141] This sanitizing agent generates peracetic acid in an amount of about 5% by weight of peracetic acid with respect to the total weight of the sanitizing agent.

[0142] More particularly, the sanitizing composition SC2 is obtained by mixing 2 percent by volume of the above-mentioned NEO-PURE sanitizing agent with 5 percent by volume of hydrogen peroxide (35%) and 97 percent by volume of water to give the following sanitizing composition SC2. As an example, for providing 100 litres of the sanitizing composition SC2, 2 litres of NEO PURE are mixed with 5 litres of hydrogen peroxide (35%) and 93 litres of water.

[0143] This sanitizing composition is particularly adapted for spraying on seeds.

[0144] Then, 1 and 1.225 kg of sunflower samples (i.e. sunflower seeds without skin, that is sunflower kernels) were inoculated with 30 or 36.75 mL of a ON TSB culture, then immediately dried at 40 C., with a fan for 12 min to return the moisture content (MC) back to untreated control (UTC).

[0145] The same day (Day one) as inoculation, the 1 kg samples were heated at 71.1 C., 82.2 C. and 93.3 C.

[0146] The following day (Day two), the 1.225 kg samples were sprayed on at rate of 60 L/t with the sanitizing composition SC2, and then heated/dried at 71.1 C., 82.2 C. or 93.3 C.

[0147] The third day (Day three), the 1.225 kg samples were sprayed on with 60 L/t dH.sub.2O (distilled water) and then heated/dried immediately at 71.1 C., 82.2 C. and 93.3 C.

[0148] Then, 45 g samples were shaken by hand in 90 mL insta Bag BPW and 1 mL was plated across 3 plates for 0 dilutions, 0.1 mL for higher dilutions on one plate.

Data and Results

[0149] The same inoculum was used at days one, two and three. Additional information concerning the inoculum are provided in the following table 5:

TABLE-US-00007 TABLE 5 Pure Inoculum Replicate 7 CFU/mL Log CFU/mL Average 1 79 790000000 8.89762709 8.89762709

Day One

[0150] Concerning the experimentation carried out at day one, the following data and results were obtained following the inoculation of the sunflower samples (see tables 6 and 7).

TABLE-US-00008 TABLE 6 UTCD1 Sample 4 CFU/g Log Count Average SD 1 245 4,900,000 6.69 6.61 0.08 2 238 4,760,000 6.68 3 177 3,540,000 6.55 4 217 4,340,000 6.64 5 157 3,140,000 6.53

TABLE-US-00009 TABLE 7 Sample MC a.sub.w Time(min) UTC 4.525 0.3833 160 F. 3.075 0.1889 30 180 F. 2.625 0.1432 30 200 F. 1.95 0.0939 30

[0151] Then, sunflower samples of day one were heated/dried in a fluid bed dryer (Sherwood Scientific Model 501 Fluid Bed Dryer), at 71.1 C., 82.2 C. and 93.3 C. The following data and results were obtained (see Tables 8, 9, 10, 11, 12, 13).

TABLE-US-00010 TABLE 8 Heated/Dried at 71.1 C. Log Sample 4 CFU/g Log Count Average SD Reduction 1 144 2,880,000 6.46 6.41 0.04 0.20 2 135 2,700,000 6.43 3 110 2,200,000 6.34 4 126 2,520,000 6.40 5 134 2,680,000 6.43

[0152] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD/2 in the following table) was set to 73.50 C. for the 71.1 C. target according to the following table 9.

TABLE-US-00011 TABLE 9 Time (min) FBD #2 Thermocoupler ( C.) 00:00 73.4 71.1 C. 00:30 73.3 71.17 01:00 73.4 71.06 01:30 73.3 71.17 02:00 73.5 71.22

TABLE-US-00012 TABLE 10 Heated/Dried at 82.2 C. Log Sample 4 CFU/g Log Count Average SD Reduction 1 98 1,960,000 6.29 6.29 0.07 0.32 2 91 1,820,000 6.26 3 98 1,960,000 6.29 4 125 2,500,000 6.40 5 81 1,620,000 6.21

[0153] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD/2 in the following table) was set to 84 C. for the 82.2 C. target according to the following table 11).

TABLE-US-00013 TABLE 11 Time (min) FBD #2 Thermocoupler( C.) 00:00 83.3 81.61 00:30 84.2 81.83 01:00 84 81.78 01:30 83.3 81.67 02:00 84.1 81.50

TABLE-US-00014 TABLE 12 Heated/Dried at 93.3 C. Log Sample 4 CFU/g Log Count Average SD Reduction 1 42 840,000 5.92 6.01 0.07 0.60 2 60 1,200,000 6.08 3 55 1,100,000 6.04 4 55 1,100,000 6.04 5 44 880,000 5.94

[0154] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD #2 in the following table) was set to 99 C. for the 93.3 C. target according to the following table 13.

TABLE-US-00015 TABLE 13 Time (min) FBD #2 Thermocoupler ( C.) 00:00 98.3 92.72 00:30 99.1 93.11 01:00 99.5 93.50 01:30 98.7 93.11 02:00 99.3 93.17

Day Two

[0155] At day two, the following data and results were obtained (see tables 14 and 15) following the inoculation.

TABLE-US-00016 TABLE 14 UTCD2 Sample 4 CFU/g Log Count Average SD 1 144 2,880,000 6.46 6.55 0.13 2 136 2,720,000 6.43 3 157 3,140,000 6.50 4 224 4,480,000 6.65 5 265 5,300,000 6.72

TABLE-US-00017 TABLE 15 Sample MC a.sub.w Time(min) UTC 4.675 0.4146 71.1 C. 3.575 0.2586 30 82.2 C. 3 0.1942 30 93.3 C. 2.325 0.1106 30

[0156] Then, the sanitizing solution SC2 was applied (i.e. sprayed) on the sunflower samples. Concerning the experimentation carried out at day two, the following data and results were obtained (i.e. spraying of the sanitizing solution SC2 (hereinafter identified as SC2 in the tables) at a rate of 60 L/t, and then heating/drying at 71.1 C., 82.2 C. and 93.3 C.), the results illustrated in the following tables 16 to 24 were obtained.

[0157] Concerning the sunflower samples sprayed with the sanitizing solution SC2 at a rate of 60 L/t and then heated/dried at 71.1 C. (see tables 16 to 18).

TABLE-US-00018 TABLE 16 60 L/t SC2 Log Sample 2 CFU/g Log Count Average SD Reduction 1 13 2,600 3.41 3.75 0.23 2.80 2 27 5,400 3.73 3 24 4,800 3.68 4 52 10,400 4.02 5 40 8,000 3.90

TABLE-US-00019 TABLE 17 60 L/t SC2 + Heating/Drying at 71.1 C. Log Total Reduction Sam- Log Aver- Log Due to ple 1 CFU/g Count age SD Reduction Drying 1 40 800 2.90 3.15 0.19 3.41 0.60 2 55 1,100 3.04 3 70 1,400 3.15 4 82 1,640 3.21 5 132 2,640 3.42

[0158] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD #2 in the following table) was set to 73 C. for the 71.1 C. target according to the following table 18.

TABLE-US-00020 TABLE 18 Time (min) FBD #2 Thermocoupler( C. 00:00 72.4 69.89 00:30 73.3 70.61 01:00 73.2 70.50 01:30 73.8 70.28 02:00 73.2 70.44

[0159] Concerning the sunflower samples sprayed with the sanitizing solution SC2 at a rate of 60 L/t and then heating/drying at 82.2 C. (see tables 19 to 21).

TABLE-US-00021 TABLE 19 60 L/t SC2 Log Sample 2 CFU/g Log Count Average SD Reduction 1 33 6,600 3.82 4.04 0.18 2.52 2 50 10,000 4.00 3 62 12,400 4.09 4 47 9,400 3.97 5 100 20,000 4.30

TABLE-US-00022 TABLE 20 60 L/t SC2 + Heating/Drying at 82.2 C. Log Total Reduction Sam- Log Aver- Log Due to ple 0 CFLI/g Count age SD Reduction Drying 1 176 352 2.55 2.75 0.19 3.81 1.29 2 186 372 2.37 3 465 930 2.97 4 280 560 2.75 5 394 788 2.90

[0160] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD #2 in the following table) was set to 82 C. for the 82.2 C. target according to the following table 21.

TABLE-US-00023 TABLE 21 Time (min) FBD #2 Thermocoupler ( C.) 00:00 82.8 82.11 00:30 81.8 81.11 01:00 81.6 80.78 01:30 82 81.11 02:00 82 80.78

[0161] Concerning the sunflower samples sprayed with the sanitizing solution SC1 at a rate of 60 L/t, and then heating/drying at 82.2 C. (see tables 22 to 24).

TABLE-US-00024 TABLE 22 60 L/t SC2 Log Sample 2 CFU/g Log Count Average SD Reduction 1 31 6,200 3.79 3.93 0.17 2.62 2 41 8,200 3.91 3 35 7,000 3.85 4 40 8,000 3.90 5 83 16,600 4.22

TABLE-US-00025 TABLE 23 60 L/t SC2 + Heating/Drying at 93.3 C. Log Total Reduction Sam- Log Aver- Log Due to ple 0 CFU/g Count age SD Reduction Drying 1 34 68 1.83 2.32 0.33 4.23 1.61 2 254 508 2.71 3 86 172 2.24 4 167 334 2.52 5 104 208 2.32

[0162] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD #2 in the following table) was set to 98.5 C. for the 93.3 C. target according to the following table 24.

TABLE-US-00026 TABLE 24 Time (min) FBD #2 Thermocoupler( C.) 00:00 97.4 92.50 00:30 98.7 92.89 01:00 98.3 93 01:30 98.2 92.72 02:00 98.8 93.17

Day Three

[0163] Concerning the experimentation carried out at day three, the following data and results were obtained. More particularly, sunflower samples were sprayed with distilled water at a rate of 60 L/t, and then heated/dried at 71.1 C., 82.2 C. and 93.3 C.), the results illustrated in the following tables 25 to 36 were obtained.

[0164] At day three the following data and results were obtained (see tables 25 and 26) following the inoculation,

TABLE-US-00027 TABLE 25 UTCD3 Sample 4 CFU/g Log Count Average SD 1 165 3,300,000 6.52 6.48 0.05 2 172 3,440,000 6.54 3 133 2,660,000 6.42 4 146 2,920,000 6.47 5 137 2,740,000 6.44

TABLE-US-00028 TABLE 26 Sample MC a.sub.w Time(min) UTC 4.625 0.4022 N/A 71.1 C. 3.65 0.2785 30 82.2 C. 3.225 0.2226 30 93.3 C. 2.825 0.2131 30

[0165] Concerning the sunflower samples sprayed with water at a rate of 60 L/t, and then heating/drying at 71.1 C. F, the following results were obtained (see tables 27 to 29)

TABLE-US-00029 TABLE 27 60 L/t dH.sub.2O Log Sample 4 CFU/g Log Count Average SD Reduction 1 165 3,300,000 6.52 6.61 0.06 0.13 2 192 3,840,000 6.58 3 214 4,280,000 6.63 4 225 4,500,000 6.65 5 221 4,420,000 6.65

TABLE-US-00030 TABLE 28 60 L/t dH2O + Heating/Drying at 71.1 C. Log Total Reduction Sam- Log Aver- Log Due to ple 3 CFU/g Count age SD Reduction Drying 1 147 294,000 5.47 5.47 0.10 1.01 1.01 2 180 360,000 5.56 3 188 376,000 5.58 4 132 264,000 5.42 5 104 208,000 5.32

[0166] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD #2 in the following table) was set to 73 C. for the 71.1 C. target according to the following table 29.

TABLE-US-00031 TABLE 29 Time (min) FBD#2 Thermocoupler ( C.) 00:00 72.8 70.22 00:30 72.8 70.27 01:00 73 70.44 01:30 72.9 70.44 02:00 73 70.61

[0167] Concerning the sunflower samples sprayed with distilled water at a rate of 60 L/t, and then heating/drying at 82.2 C., the following results were obtained (see Tables 30 and 32).

TABLE-US-00032 TABLE 30 60 L/t dH.sub.20 Log Log Sample 4 CFU/g Count Average SD Reduction 1 181 3,620,000 6.56 6.51 0.05 0.04 2 181 3,620,000 6.56 3 156 3,120,000 6.49 4 139 2,780,000 6.44 5 163 3,260,000 6.51

TABLE-US-00033 TABLE 31 60 L/t dH.sub.20 + Heating/Drying at 82.2 C. Log Total Reduction Sam- Log Aver- Log Due to ple 3 CFU/g Count age SD Reduction Drying 1 66 132,000 5.12 5.18 0.12 1.30 1.30 2 50 100,000 5.00 3 94 188,000 5.27 4 92 184,000 5.26 5 88 176,000 5.25

[0168] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD #2 in the following table) was set to 82.5 C. for the 71.1 C. target according to the following table 32.

TABLE-US-00034 TABLE 32 Time (min) FBD #2 Thermocoupler ( C.) 00:00 82.7 81.28 00:30 82.6 81.11 01:00 82.8 81.17 01:30 82.5 80.56 02:00 82.7 81.67

[0169] Concerning the sunflower samples sprayed with distilled water at a rate of 60 L/t, and then heated/dried at 93.3 C., the following results were obtained (see Tables 33 and 35).

TABLE-US-00035 TABLE 33 60 L/t dH.sub.20 Log Log Sample 4 CFU/g Count Average SD Reduction 1 164 3,280,000 6.52 6.43 0.07 0.05 2 150 3,000,000 6.48 3 127 2,540,000 6.40 4 114 2,280,000 6.36 5 122 2,440,000 6.39

TABLE-US-00036 TABLE 34 60 L/t dH.sub.2O + Heating/Drying at 93.3 C. Log Total Reduction Sam- Log Aver- Log Due to ple 3 CFU/g Count age SD Reduction Drying 1 43 86,000 4.93 4.95 0.07 1.53 1.48 2 39 78,000 4.89 3 41 82,000 4.91 4 58 116,000 5.06 5 44 88,000 4.94

[0170] The fan of the Sherwood Scientific Model 501 Fluid Bed Dryer (FBD #2 in the following table) was set to 98.000 for the 93.3 C. target according to the following table 35.

TABLE-US-00037 TABLE 35 Time (min) FBD #2 Thermocoupler ( C.) 00:00 98.2 93.28 00:30 97.5 92.72 01:00 98.3 93.56 01:30 97.6 93.22 02:00 98 93.56

TABLE-US-00038 TABLE 36 Avg. Avg. SAMPLE UTC Treated Log Treated + Dried Log Log red. UTC's (rate, Log S Log S red. Log Std red. due to % UTC's drying T) CFU/g dev CFU/g dev CFU/g CFU/g dev CFU/g drying M.C. a.sub.w SC2, 6.55 0.13 3.75 0.23 2.80 3.15 0.19 3.41 0.60 4.68 0.41 60 L/t, 71.1 C. SC2, 4.04 0.18 2.52 2.75 0.19 3.81 1.29 60 L/t, 82.2 C. SC2, 3.93 0.17 2.62 2.32 0.33 4.23 1.61 60 L/t, 93.3 C. Water, 6.48 0.05 6.61 0.06 0.13 5.47 0.10 1.01 1.01 4.63 0.40 60 L/t, 71.1 C. Water, 6.51 0.05 0.04 5.18 0.12 1.30 1.30 60 L/t, 82.2 C. Water, 6.43 0.07 0.05 4.95 0.07 1.53 1.48 60 L/t, 93.3 C. No 6.61 0.08 N/A N/A N/A 6.41 0.04 0.20 0.20 4.53 0.38 treatment, 71.1 C. No 6.29 0.07 0.32 0.32 treatment, 82.2 C. No 6.01 0.07 0.60 0.60 treatment, 93.3 C.

[0171] Table 36 summarizes the efficacy results on reduction of E. faecium in sunflower kernels when treated with either sanitizing composition SC2 or water at a rate of 60 L/t, or no treatment and after drying at three different temperatures (71.1 C., 82.2 C. and 93.3 C.).

[0172] When the inoculated sunflower kernels were treated with sanitizing composition SC2, and heated/dried at 71.1 C., 82.2 C., and 93.3 c., the log reduction achieved on E. faecium was 3.41, 3.81, and 4.23, respectively.

[0173] However, when the sunflower kernels were treated with water, less than 1.5 log reduction was achieved, even at the highest temperature of 93.3 C.

[0174] Furthermore, when the sunflower kernels were not treated with any liquid, but heated/dried at the three temperatures, the least efficacy on E. faecium was observed, ranging from 0.2 to 0.60 log CFU/g reduction.

[0175] In addition, when the theoretical value of the log reduction of the sanitizing composition SC2 and temperature alone (SC2 (60 L/t)+temperature (theoretical)) was calculated, the results were 0.4 to 1.01 log CFU/g lower than the values achieved when the sanitizing composition SC2 and temperature were actually combined in the treatment of sunflower kernels, with an increase in the synergistic effect observed as drying temperature increased.

CONCLUSION

[0176] It can be concluded that there is a synergistic effect between the sanitizing composition SC2 and the heating/drying treatment applied during the heating/drying process at different temperatures (71.1 C., 82.2 C. and 93.3 C.) on reduction of E. faecium in sunflower kernels.

Example 3

[0177] Example 2 was repeated with samples of almonds, except: [0178] Almonds were treated (sprayed) with a sanitizing composition SC3 (hereinafter called SC3) resulting from the admixture of 2% by volume of the Neo Pure sanitizing agent (defined in example 1), 7% by volume of hydrogen peroxide (35%) and 91% by volume of water, at a rate of 100 L of SC3 per ton of almonds, [0179] Almonds were treated (sprayed) with water at a rate of 100 L per ton of almonds, or [0180] Almonds were untreated; and
then heated/dried for 10 minutes at 104.4 C. in either a lab dryer or in a continuous drier (e.g. a fluid bed dryer of the type Sherwood Scientific Model 501 Fluid Bed Dryer). Results are reported in the following tables 36 to 51.

[0181] Innoculation of almonds is similar to the one carried out with sunflower kernels in example 2.

Replicate One

[0182] Tables 36 to 38 represent data concerning untreated samples, samples treated with SC3 and samples treated with SC3 and then heated/dried at 104.4 C. in a lab heating/dryer oven.

TABLE-US-00039 TABLE 37 UTC Average Stdev Log10 Log10 Log 10 Sample 4 CFU/g CFU/g CFU/g CFU/g 1 65 1.30E+06 6.11 6.04 0.05 2 54 1.08E+06 6.03 3 53 1.06E+06 6.03 4 56 1.12E+06 6.05 5 48 9.60E+05 5.98

TABLE-US-00040 TABLE 38 TREATED WITH SC3 Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 2 CFU/g CFU/g CFU/g CFU/g red sample % MC aw 1 80 1.60E+04 4.20 3.94 0.31 2.10 UTC 5.3 0.56 2 61 1.22E+04 4.09 1.78* T + D 4.75 0.52 3 51 1.02E+04 4.01 4 49 9.80E+03 3.99 5 13 2.60E+03 3.41 *minimum log reduction

TABLE-US-00041 TABLE 39 TREATED WITH SC3 AND THEN HEATED/DRIED FOR 10 MIN Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 0 CFU/g CFU/g CFU/g CFU/g red 1 21 4.20E+01 1.62 1.97 0.20 4.07 2 50 1.00E+02 2.00 3.86* 3 67 1.34E+02 2.13 4 54 1.08E+02 2.03 5 58 1.16E+02 2.06 *minimum log reduction

[0183] Tables 40 to 42 represent data concerning untreated samples, samples treated with distilled water, and samples treated with distilled water and then heated/dried at 104.4 C. in a lab heating/dryer oven.

TABLE-US-00042 TABLE 40 UTC Average Stdev Log10 Log10 Log 10 Sample 4 CFU/g CFU/g CFU/g CFU/g 1 65 1.30E+06 6.11 6.04 0.05 2 54 1.08E+06 6.03 3 53 1.06E+06 6.03 4 56 1.12E+06 6.05 5 48 9.60E+05 5.98

TABLE-US-00043 TABLE 41 TREATED WITH DISTILLED WATER Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 3 CFU/g CFU/g CFU/g CFU/g red sample % MC a.sub.w 1 592 1.18E+06 6.07 6.04 0.05 0.00 UTC 5.3 0.56 2 486 9.72E+05 5.99 T + D 4.9 0.5 3 640 1.28E+06 6.11 4 480 9.60E+05 5.98 5 560 1.12E+06 6.05

TABLE-US-00044 TABLE 42 TREATED WITH DISTILLED WATER AND THEN HEATED/DRIED FOR 10 MIN Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 3 CFU/g CFU/g CFU/g CFU/g red 1 48 9.60E+04 4.98 4.87 0.10 1.17 2 44 8.80E+04 4.94 1.04* 3 28 5.60E+04 4.75 4 39 7.80E+04 4.89 5 32 6.40E+04 4.81 *minimum log reduction

[0184] Tables 43 and 44 represent data concerning untreated samples, and untreated samples heated/dried at 104.400 in a lab heating/dryer oven.

TABLE-US-00045 TABLE 43 UTC Average Stdev Log10 Log10 Log 10 Sample 4 CFU/g CFU/g CFU/g CFU/g 1 65 1.30E+06 6.11 6.04 0.05 2 54 1.08E+06 6.03 3 53 1.06E+06 6.03 4 56 1.12E+06 6.05 5 48 9.60E+05 5.98

TABLE-US-00046 TABLE 44 HEATED/DRIED FOR 10 MIN DRIED FOR 10 MIN Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 3 CFU/g CFU/g CFU/g CFU/g red sample % MC a.sub.w 1 225 4.50E+05 5.65 5.72 0.06 0.32 UTC 5.3 0.56 2 227 4.54E+05 5.66 0.25* T + D 4.55 0.5 3 297 5.94E+05 5.77 4 280 5.60E+05 5.75 5 305 6.10E+05 5.79 *minimum log reduction

Replicate Two

[0185] Tables 45 to 47 represent data concerning untreated samples, samples treated with SC3 and samples treated with SC3 and then heated/dried at 104.400 in a fluid bed dryer of the type Sherwood Scientific Model 501 Fluid Bed Dryer)

TABLE-US-00047 TABLE 45 UTC Average Stdev Log10 Log 10 Log 10 Sample 4 CFU/g CFU/g CFU/g CFU/g 1 31 6.20E+05 5.79 5.94 0.10 2 39 7.80E+05 5.89 3 44 8.80E+05 5.94 4 56 1.12E+06 6.05 5 50 1.00E+06 6.00

TABLE-US-00048 TABLE 46 TREATED WITH SC3 Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 2 CFU/g CFU/g CFU/g CFU/g red sample % MC a.sub.w 1 43 8.60E+03 3.93 3.98 0.04 1.96 UTC 5.2 0.57 2 55 1.10E+04 4.04 1.79* T + D 4.6 0.51 3 48 9.60E+03 3.98 4 50 1.00E+04 4.00 5 44 8.80E+03 3.94 *minimum log reduction

TABLE-US-00049 TABLE 47 TREATED WITH SC3 AND THEN HEATED/DRIED FOR 10 MIN Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 0 CFU/g CFU/g CFU/g CFU/g red 1 3 6.00E+00 0.78 0.94 0.29 5.00 2 14 2.80E+01 1.45 4.35* 3 3 6.00E+00 0.78 4 3 6.00E+00 0.78 5 4 8.00E+00 0.90 *minimum log reduction

[0186] Tables 48 to 50 represent data concerning untreated samples, samples treated with distilled water, and samples treated with distilled water and then heated/dried at 104.400 in a fluid bed dryer of the type Sherwood Scientific Model 501 Fluid Bed Dryer)

TABLE-US-00050 TABLE 48 UTC Average Stdev Log10 Log10 Log 10 Sample 4 CFU/g CFU/g CFU/g CFU/g 1 31 6.20E+05 5.79 5.94 0.10 2 39 7.80E+05 5.89 3 44 8.80E+05 5.94 4 56 1.12E+06 6.05 5 50 1.00E+06 6.00

TABLE-US-00051 TABLE 49 TREATED WITH DISTILLED WATER Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 3 CFU/g CFU/g CFU/g CFU/g red sample % MC a.sub.w 1 99 1.98E+05 5.30 5.18 0.09 0.75 UTC 5.2 0.57 2 64 1.28E+05 5.11 0.50* T + D 5.05 0.55 3 80 1.60E+05 5.20 4 59 1.18E+05 5.07 5 87 1.74E+05 5.24 *minimum log reduction

TABLE-US-00052 TABLE 50 TREATED WITH DISTILLED WATER AND THEN HEATED/DRIED FOR 10 MIN TREATED + HEATED/DRIED Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 2 CFU/g CFU/g CFU/g CFU/g red 1 129 2.58E+04 4.41 4.56 0.11 1.37 2 230 4.60E+04 4.66 1.12* 3 180 3.60E+04 4.56 4 158 3.16E+04 4.50 5 237 4.74E+04 4.68 *minimum log reduction

[0187] Tables 51 and 52 represent data concerning untreated samples, samples heated/dried at 104.400 in a fluid bed dryer of the type Sherwood Scientific Model 501 Fluid Bed Dryer)

TABLE-US-00053 TABLE 51 UTC Average Stdev Log10 Log10 Log 10 Sample 4 CFU/g CFU/g CFU/g CFU/g 1 31 6.20E+05 5.79 5.94 0.10 2 39 7.80E+05 5.89 3 44 8.80E+05 5.94 4 56 1.12E+06 6.05 5 50 1.00E+06 6.00

TABLE-US-00054 TABLE 52 HEATED/DRIED FOR 10 MIN Average Stdev Log Log10 Log10 Log 10 CFU/g Sample 3 CFU/g CFU/g CFU/g CFU/g red sample % MC a.sub.w 1 168 3.36E+05 5.53 5.48 0.07 0.45 UTC 5.2 0.57 2 184 3.68E+05 5.57 0.23* T + H/D 4.45 0.5 3 134 2.68E+05 5.43 4 126 2.52E+05 5.40 5 155 3.10E+05 5.49 *minimum log reduction

[0188] Comparison between replicates one and two

TABLE-US-00055 TABLE 53 Average log reduction of Replicate one and two Log reduction Log reduction after treatment after drying (minimum) (minimum) MC % UTC N/A N/A 5.25 SC3 treated 1.79 4.11 4.7 Water treated 0.25 1.08 5 Just dried 0 0.24 4.5

Demonstration of a Synergistic Effect

[0189] Therefore, the above-mentioned results appearing in table 53 can be summarized as illustrated in the following table 54:

TABLE-US-00056 TABLE 54 Treatment Log reduction achieved Heat only 0.24 Water only 0.25 Water + heating/drying 1.08 SC3 only 1.79 SC3 + heating/drying 4.11 (instead of 1.79 + 0.24 = 2.03; i.e. 2.08 more log reduction is achieved due to synergistic effect)

[0190] This table clearly illustrate that the SC3 followed by a heating/drying step showed a synergistic effect. Without being bound to the theory, the Applicant believes that [0191] the synergistic effect is probably achieved due to the fact that the sanitizing composition gets vaporized during the heating/drying step and continues to decontaminate the food during this step; and [0192] that the effect of the sanitizing composition gets enhanced as its temperature goes up during the drying stage.

[0193] Thus, the Applicant believes that this synergistic effect can happen on any type of food, regardless of the type of seeds, including the preservation of the viability of said seeds.

[0194] The above description of the embodiments should not be interpreted in a limiting manner since other variations, modifications and refinements are possible within the scope of the present invention. Accordingly, it should be understood that various features and aspects of the disclosed embodiments can be combined with or substituted for one another in order to form varying modes of the disclosed invention. The scope of the invention is defined in the appended claims and their equivalents.