Use of a chestnut tannin extract as acaricidal agent

Abstract

The use is described of a chestnut tannin extract (Castanea sativa Mill.), as a non-phytotoxic acaricidal agent, alone or in a mixture with other active principles having an acaricidal activity.

Claims

1. A method of controlling, limiting, rejecting or destroying arachnids comprising applying a chestnut tannin extract (Castanea sativa Mill.), as a non-phytotoxic acaricidal agent, to an agricultural crop, to soil or by seed tanning, alone or in a mixture with other active ingredients having an acaricidal activity.

2. The method according to claim 1, comprising obtaining the chestnut tannin extract by means of a process, without the use of chemical additives, which comprises an extraction step in water by leaching starting from a vegetable biomass, filtering, and concentrating with physical means.

3. The method according to claim 2, wherein the physical means are nanofiltration and/or reverse osmosis.

4. The method according to claim 2, wherein the chestnut tannin extract is in liquid or solid form.

5. The method according to claim 2 wherein the vegetable biomass comprises any part of the chestnut plant selected from the group consisting of trunk, leaves, branches or bark or any combination thereof.

6. The method according to claim 1 comprising applying the chestnut tannin extract without other active ingredients.

7. The method according to claim 1 wherein the chestnut tannin extract comprises a fraction corresponding to a concentrate obtained from a first nanofiltration step, a fraction corresponding to a concentrate obtained from a second nanofiltration step, a fraction corresponding to a concentrate obtained from a reverse osmosis step, or a fraction corresponding to a powder obtained at the end of a drying step, alone or in any combination thereof.

8. The method according to claim 1 comprising applying the chestnut tannin extract, alone or in a mixture with other active ingredients having an acaricidal activity, in the liquid or solid state to agricultural crops for prevention or treatment.

9. The method according to claim 1, comprising applying the chestnut tannin extract in a quantity ranging from 100 g to 10,000 g per hectare of agricultural crop.

10. A non-phytotoxic agronomic acaricidal formulation, comprising chestnut tannin extract present in a quantity ranging from 90 to 95% by weight, one or more surfactants present in a quantity ranging from 2 to 6% by weight, one or more dispersants present in a quantity ranging from 2 to 6% by weight and an anti-foam agent present in a quantity ranging from 1 to 2% by weight, all percentages based on the total weight of the formulation, wherein the chestnut tannin extract is obtained by means of a process, without the use of chemical additives, which comprises an extraction step in water by leaching, starting from a vegetable biomass, filtering, and concentrating with physical means.

11. The formulation according to claim 10, wherein the chestnut tannin extract is chestnut tannin extract in powder form mixed with a surfactant based on sodium alkyl sulfonate, a dispersant based on alkyl naphthalene sulfonates and a chemically and agronomically acceptable anti-foam agent.

12. The formulation according to claim 10, wherein the chestnut tannin extract is in powder form and the surfactant is a wetting agent.

13. The formulation according to claim 10, wherein said formulation also comprises one or more additional active ingredients.

14. A method for controlling mites comprising applying the formulation according to claim 11 to an agricultural crop, to soil or by seed tanning.

15. The method of claim 14, comprising applying the formulation to an agricultural crop, in the liquid or solid state, for prevention or treatment.

16. The method of claim 2 wherein the process further comprises a final drying step.

17. The method of claim 4 wherein the chestnut tannin extract is in solid form.

18. The method of claim 9 comprising applying the chestnut tannin extract in a quantity ranging from 500 g to 8,000 g per hectare of agricultural crop.

19. The formulation of claim 10 wherein the process further comprises a final drying step.

20. The formulation of claim 13, wherein at least one of the one or more additional active ingredients have an acaricidal activity.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is an exemplary chromatographic profile of the chestnut tannin extract of Fraction 5 obtained by nanofiltration concentration.

(2) FIG. 2 is an exemplary chromatographic profile of the chestnut tannin extract of Fraction 8 obtained by spray drying and subsequent re-dispersing in water.

(3) FIG. 3 is a comparison between the chestnut tannin extract obtained by Example 2 and the chestnut tannin extract of Fraction 8.

(4) FIG. 4 depicts the results of a field test for determination of the acaricidal activity of the formulation of Example 2 against the eggs of Tetranychus urticae on aubergines.

(5) FIG. 5 depicts the results of a field test for determination of the acaricidal activity of the formulation of Example 2 against nymphs of Tetranychus urticae on aubergines.

(6) FIG. 6 depicts the results of a field test for determination of the acaricidal activity of the formulation of Example 2 against adults of Tetranychus urticae on aubergines.

(7) FIG. 7 depicts the results of a field test for determination of the acaricidal activity of the formulation of Example 2 against the total mobile forms of Tetranychus urticae on aubergines.

(8) FIG. 8 depicts the results of a field test for determination of the acaricidal activity of the formulation of Example 2 against the eggs of Tetranychus sp. on soybeans.

(9) FIG. 9 depicts the results of a field test for determination of the acaricidal activity of the formulation of Example 2 against mobile forms of Tetranychus sp. on soybeans.

(10) FIG. 10 depicts of the results of a field test for determination of the acaricidal activity of the formulation of Example 2 against the total mobile forms of Tetranychus sp. on soybeans.

EXAMPLES

(11) The following examples are provided for purely illustrative purposes of the present invention and should not be considered as limiting the protection scope, as defined by the enclosed claims.

Example 1

(12) Preparation and Production of a Tannin Extract

(13) Applying the aforementioned method of hot aqueous extraction (leaching) of chestnut wood and cold fractionation using the membrane technology, 6,000 kg of chestnut wood were treated with 20,000 kg of fresh water and the following eight fractions were obtained and analyzed: 1. Filtered tannic broths; 2. Nanofiltration permeate I; 3. Nanofiltration concentrate I; 4. Nanofiltration permeate II; 5. Nanofiltration concentrate II; 6. Osmosis permeate; 7. Osmosis concentrate; 8. Powder.

(14) A Luna C18 column of 2504.60 mm, 5 m (Phenomenex, Torrance, Calif.), was used for the HPLC analysis of the single fractions, the mobile phase used consisted of H.sub.2O (pH 3.2 for HCOOH), (A) and CH.sub.3CN (B). A four-ramp linear gradient was applied, a flow-rate of 0.8 ml/min, for 55 minutes. The elution profile used is the following: initially 100% A, the solvent A was then brought to 85% in 20 minutes, kept constant for 5 minutes, reduced to 75% in 10 minutes, kept constant for 8 minutes, finally brought to 0% (100% B) in 5 minutes and kept constant for 4 minutes, then returning to the initial conditions in 3 minutes. The gallic derivatives were calibrated at 280 nm with gallic acid, the ellagic derivatives at 254 nm with ellagic acid.

(15) The chromatographic profiles relating to two fractions are provided by way of example: the first, liquid, obtained by nanofiltration concentration (Fraction 5.), having a density at 20 C. of 1.30 g/ml (FIG. 1), and the second obtained by spray drying starting from the latter (Fraction 8, FIG. 2) and subsequently re-dispersed in water.

(16) As previously indicated, the fractions that can be used and are used for the purposes of the present invention are the following: Fraction 3. Nanofiltration concentrate I Fraction 5. Nanofiltration concentrate II Fraction 7. Osmosis concentrate Fraction 8. Powder

(17) respectively characterized by the following contents of chestnut tannin extract: Fraction 3, having a content of 18% weight/weight; Fraction 5, having a content of 50% weight/weight; Fraction 7, having a content of 15% weight/weight; Fraction 8, having a content of 100% weight/weight.

(18) As indicated above, content of chestnut tannin extract refers to the sum of the concentrations, expressed in weight %, of derivatives of gallic acid, pure gallic acid, derivatives of ellagic acid, pure ellagic acid and all other components not belonging to these four categories, i.e.: pentosans, hexosans, oligosaccharides, hemicellulose, simple organic acids and inorganic salts, with respect to the total weight of the solution, in the case of a liquid fraction, or with respect to the total weight of the solid, in the case of powder.

Example 2

(19) Preparation of a Formulation of Tannin Extract in Powder Form with Improved Re-Dispersibility

(20) The tannin extract in powder form (Fraction 8.) with a tannin content equal to 100% by weight obtained in Example 1, was mixed with the following products in the proportions indicated hereunder:

(21) TABLE-US-00001 weight % Fraction 8. 91 Wetting agent based on sodium ligninsulfonate (Bretax S) 5 Anti-foam agent based on a mixture of organic compounds 1.5 and mineral oil absorbed on an inorganic carrier, whose main component is benzene, mono-C10-13-alkyl-derivative, distillation residues (Defomex AP/188) Dispersing agent based on sodium naphthalene sulfonate 2.5 (Madeol AG/W90)

(22) The powder obtained has the same chemical characteristics as Fraction 8. (main component), but is characterized by a much higher re-dispersion capacity/property in water.

(23) In order to demonstrate this, Fraction 8. and the formulation described above were taken and re-dispersed in water.

(24) FIG. 3 shows how the formulated powder (on the left, Formulation Example 2) has a better re-dispersion than the original powder (on the right, Fraction 8.)).

(25) More specifically, a parallel experiment was carried out in which, at a temperature of 23 C., 100 g of both products in powder form were added to 900 g of water under constant stirring. The stirring time was 3 minutes for both, at the end of which the quantity of powder remaining undissolved was evaluated: the formulated powder did not show any residues, whereas with Fraction 8. as such, 27.7 g (corresponding to 27.7%) remained undissolved, forming clots and also creating foam (as can be seen from FIG. 3).

Example 3

(26) Determination of the Acaricidal Activity of the Formulation of Example 2 Against Tetranychus urticae on Aubergines

(27) a) Activity of Tetranychus urticae on the Eggs (Expressed as % of Non-Verified Attack

(28) During the summer of 2016, a field test was carried out (locality Andria, region of Puglia) on aubergines (variety Solanum melongena).

(29) The plant was treated with the following products: Formulation of Example 2 dispersed in water and applied in a ratio of 0.5 kg/ha, dosing a volume equivalent to 1,000 l/ha; Formulation of Example 2 dispersed in water and applied in a ratio of 1 kg/ha, dosing a volume equivalent to 1,000 l/ha; Formulation of Example 2 dispersed in water and applied in a ratio of 2 kg/ha, dosing a volume equivalent to 1,000 l/ha; Formulation of Example 2 dispersed in water and applied in a ratio of 4 kg/ha, dosing a volume equivalent to 1,000 l/ha; Borneo (commercial product based on etoxazole) diluted in water in a ratio of 500 ml/1,000 liters, dosing a volume equivalent to 1,000 l/ha. The dosage applied is the dosage recommended on the label.

(30) A non-treated blank, i.e. a block of land not treated with any product, was then evaluated for comparative purposes.

(31) Each block of land measured 115 m.sup.2. For each treatment provided (a total of 6 including the non-treated block), 4 replications were effected, for a total of 24 blocks treated, following a randomized on-field treatment scheme of the blocks (RCB: Randomized Complete Block).

(32) In the test, the formulation of Example 2 was tested, at different doses, and compared with a known anti-mite commercial product, more specifically the product Borneo, against infestation by Tetranychus urticae, also known as Red spider, a mite well-known in the agricultural field, whose damage is extremely feared and conspicuous.

(33) The applications were effected on June 25, July 5 and July 15 using a spray system directly on the leaves. The products were distributed with a dosage equal to 1,000 liters/ha, a total of 1,000 liters:10,000 m.sup.2(154 m.sup.2)=6 liters were then sprayed for each test.

(34) The results are indicated in Table 1 and in the relative graph of FIG. 4.

(35) The controls indicated in Table 1 were effected immediately before the subsequent applications of July 5 and July 15.

(36) The percentages indicated in the Table correspond to the percentage of crop not attacked by the eggs of the mite: the percentage is equal to 100% when the crop was not attacked by the mite, whereas the percentage is equal to 0% when the whole crop was attacked by the mite.

(37) The evaluation was effected on a sample of 25 leaves per block.

(38) TABLE-US-00002 TABLE 1 Effectiveness of the Formulation of Example 2 on the eggs of Tetranychus urticae on aubergines 05-July 15-July Days after 1.sup.st application (DAA) (June 25) 10 20 Non-treated blank 0.00 0.00 Formulation of Example 2 52.26 51.79 0.5 Kg/ha Formulation of Example 2 48.39 50.62 1 Kg/ha Formulation of Example 2 51.61 51.98 2 Kg/ha Formulation of Example 2 60.65 56.34 4 kg/ha BORNEO 500 ml/ha 70.32 70.86

(39) It is evident from the relative graph, in FIG. 4, that the tannin extract, even at its lowest dosage, is able to inhibit about 50% of attack by the eggs of Tetranychus urticae, whereas in the blocks of untreated blanks the value obtained for both tests were equal to zero, i.e. 100% attack.

(40) A slight dosage/effect correlation can be noted with respect to the tannin extract: with an increase in the dosage, the response improves (even if not proportionally).

(41) It should be pointed out that the difference in activity between the tannin extract (at the maximum dosage tested of 4 kg/ha) and the commercial reference product (Borneo) is rather small (12% less considering the average between the two tests). It is therefore absolutely reasonable to assume that higher dosages of tannin extract can lead to completely equivalent results, if not higher, in numerical terms, to those of the commercial reference product (Borneo), with the enormous advantage of using a completely natural product instead of a synthetic chemical product.

(42) b) Activity on Tetranychus urticae Nymphs (Expressed as % of Non-Verified Attack)

(43) The test was carried out as indicated in item a) and the results of the activity of the products tested on the nymphs are shown in Table 2 and in the relative graph of FIG. 5. The percentages indicated in the table correspond to the percentages of crop not attacked by the nymphs of mites: the percentage is equal to 100% when the crop was not attacked by the mite, whereas the percentage is equal to 0% when the whole crop was attacked by the mite.

(44) The evaluation was carried out on a sample of 25 leaves per block.

(45) TABLE-US-00003 TABLE 2 Effectiveness of the Formulation of Example 2 on the nymphs of Tetranychus urticae on aubergines 05-July 15-July Days after 1.sup.st application (DAA) (June 25) 10 20 Non-treated blank 0.00 0.00 Formulation of Example 2 29.95 62.00 0.5 Kg/ha Formulation of Example 2 59.24 64.00 1 Kg/ha Formulation of Example 2 54.50 62.00 2 Kg/ha Formulation of Example 2 65.88 71.00 4 kg/ha BORNEO 500 ml/ha 72.93 75.70

(46) It is evident from the relative graph, in FIG. 5, that the chestnut tannin extract (excluding the lowest dosage, after the first 10 days of treatment) shows an activity completely comparable to that of the reference product.

(47) Also in this case, the dosage/effect correlation is not particularly marked.

(48) 20 days after the first treatment, the difference in the activity between the tannin extract (at the maximum dosage tested of 4 kg/ha, and the commercial reference product (Borneo) is minimum. It is therefore reasonable to assume that higher dosages of chestnut tannin can lead to completely equivalent results, if not higher, in numerical terms, to those of the commercial reference product (Borneo) with the enormous advantage of using a completely natural product instead of a synthetic chemical product.

(49) c) Activity on Adults of Tetranychus urticae (Expressed as % of Non-Verified Attack)

(50) The test was carried out as indicated in item a) and the results of the activity on the adults are shown in Table 3 and in the relative graph of FIG. 6. The percentages indicated in Table 3 correspond to the percentage of crop not attacked by the adult mites: the percentage is equal to 100% when the crop was not attacked by the mite, whereas the percentage is equal to 0% when the whole crop was attacked by the mite

(51) The evaluation was carried out on a sample of 25 leaves per block.

(52) TABLE-US-00004 TABLE 3 Effectiveness of the Formulation of Exmple 2 on the adults of Tetranychus urticae on aubergines 05-July 15-July Days after 1.sup.st application (DAA) (June 25) 10 20 Non-treated blank 0.00 0.00 Formulation of Example 2 7.5 49.41 0.5 Kg/ha Formulation of Example 2 18.00 51.18 1 Kg/ha Formulation of Example 2 14.50 60.42 2 Kg/ha Formulation of Example 2 38.50 64.38 4 kg/ha BORNEO 500 ml/ha 75.50 80.98

(53) It is evident from the relative graph, in FIG. 6, that the chestnut tannin extract, at the highest dosages and 20 days after the first treatment, shows an activity not far from that of the reference product. Also in this case the dosage/effect correlation is not particularly marked, but seems more linear with respect to the previous tests.

(54) The considerable increase in the activity of the chestnut tannin extract with each passing day, should be noted however, which makes it completely reasonable to assume that in the subsequent 5/10 days of observation, the results obtained with the reference product, can be surpassed.

(55) d) Activity on the Total Mobile Forms of Tetranychus urticae (Expressed as Total Number of the Mobile Forms)

(56) The test was carried out as indicated in item a) and the results of the activity on the total mobile forms are shown in Table 4 and in the relative graph of FIG. 7.

(57) The numbers indicated in Table 4 therefore correspond to the total number of mobile forms, i.e. to the number of living forms still present on the crop: the closer the value found is to zero, the more effective the treatment has been, as most of the population of mites has been eradicated.

(58) The evaluation was carried out on a sample of 25 leaves per block.

(59) TABLE-US-00005 TABLE 4 Effectiveness of the Formulation of Example 2 on the total mobile forms of Tetranychus urticae on aubergines 05-July 15-July Days after 1.sup.st application (DAA) (June 25) 10 20 Non-treated blank 141.50 141.25 Formulation of Example 2 101.75 64.00 0.5 Kg/ha Formulation of Example 2 82.50 62.50 1 Kg/ha Formulation of Example 2 85.50 58.25 2 Kg/ha Formulation of Example 2 64.00 50.00 4 kg/ha BORNEO ml/ha 38.00 33.50

(60) It is evident from the relative graph, in FIG. 7, that chestnut tannin extract, in general, allows the overall population of mites present on the crop to be controlled, giving the best results 20 days after the first treatment.

(61) A more pronounced dosage/effect correlation can be noted with respect to the previous tests, even if not linear.

(62) e) Evaluation of the General Phototoxicity Shown by the Products Tested

(63) The test was carried out as indicated in item a).

(64) The objective of this test is to evaluate and quantify any form of phytotoxicity (chlorosis, necrosis, abnormal staining, whitening, etc.) that can appear on the crop under examination following application of the products tested, 10 and 20 days after the first treatment. The values indicated in Table 5 correspond to the percentage of plant that has suffered damage (chlorosis, necrosis, abnormal staining, whitening, etc.): the closer the value found is to zero, the more the treatment has proved to be non-phytotoxic.

(65) TABLE-US-00006 TABLE 5 Evaluation of the impact of the Formulation of Example 2 on the general phytotoxicity on aubergines 05-July 15-July Days after 1.sup.st application (DAA) (June 25) 10 20 Non-treated blank 0.00 0.00 Formulation of Example 2 0.00 0.00 0.5 Kg/ha Formulation of Example 2 0.00 0.00 1 Kg/ha Formulation of Example 2 0.00 0.00 2 Kg/ha Formulation of Example 2 0.00 0.00 4 kg/ha BORNEO 500 ml/ha 0.00 0.00

(66) It is evident that the tannin extract, object of the invention, does not show any phytotoxicity with respect to the crop, at any dosage. Chlorosis, necrosis, abnormal staining, whitening, were tested.

Example 4

(67) Determination of the Acaricidal Activity of the Formulation of Example 2 Against Tetranychus sp. on soybeans

(68) a) Activity on the Eggs of Tetranychus sp. (Expressed as % of Non-Verified Attack

(69) In August 2016, a field test was carried out (locality Pettorazza Grimani, province of Rovigo, region Veneto) on soybeans (variety Glycine Max).

(70) The plant was treated with the following products: Formulation of Example 2 dispersed in water and applied in a ratio of 1 kg/ha, dosing a volume equivalent to 600 l/ha; Formulation of Example 2 dispersed in water and applied in a ratio of 2 kg/ha, dosing a volume equivalent to 600 l/ha Formulation of Example 2 dispersed in water and applied in a ratio of 4 kg/ha, dosing a volume equivalent to 600 l/ha Formulation of Example 2 dispersed in water and applied in a ratio of 8 kg/ha, dosing a volume equivalent to 600 l/ha; MATACAR FL (commercial product based on Hexythiazox) diluted in water at a ratio of 20 g/100 liters, dosing a volume equivalent to 600 l/ha. The dosage applied is that recommended in the label.

(71) A non-treated blank, i.e. a block of land not treated with any product, was also evaluated for comparative purposes.

(72) Each block of land measured 310 m2. 4 replications were effected for each treatment (6, including the non-treated block) for a total of 24 blocks tested, following a randomized on-field treatment scheme of the blocks (RCB: Randomized Complete Block).

(73) In the test, the formulation of Example 2 was tested, at different doses, and compared with a known anti-mite commercial product, more specifically the product Matacar FL, against infestation by Tetranychus sp., a mite well-known in the agricultural field, whose damage is extremely feared and conspicuous.

(74) The products were distributed with a dosage equivalent to 600 liters/ha, therefore a total of 600 liters: 10,000 m.sup.2(304 m.sup.2)=7.2 liters were sprayed for each test. The only application was effected on August 11 using a spray system directly on the leaves. On August 13, August 18 and August 25 the tests shown in the following Tables were carried out.

(75) The results of the activity on the eggs of Tetranychus sp. on soybeans are shown in Table 6 and in the relative graph of FIG. 8. The percentages indicated in Table 6 correspond to the percentages of crop which was not attacked by the eggs of the mite: if the crop was not attacked by the mite, the percentage is equal to 100%, whereas if the whole crop was attacked, the percentage is 0%.

(76) The evaluation was carried out on a sample of 25 leaves per block.

(77) TABLE-US-00007 TABLE 6 Effectiveness of the Formulation of Example 2 on eggs of Tetranychus sp. on soybeans 13-Aug. 18-Aug. 25-Aug. Days after application (DAA) (August 11) 2 7 14 Non-treated blank 0.0 0.0 0.0 Formulation of Example 2 18.1 19.7 32.9 1 Kg/ha Formulation of Example 2 13.6 9.8 22.5 2 Kg/ha Formulation of Example 2 27.4 50.0 25.8 4 kg/ha Formulation of Example 2 46.6 52.1 55.9 8 kg/ha MATACAR FL 120 g/ha 31.5 43.4 52.2

(78) It is evident from the relative graph, in FIG. 8, that the chestnut tannin extract (already at a dosage of 4 kg/ha) shows a completely competitive activity with respect to the reference product, even as far as surpassing its effectiveness at the maximum dosage in all the tests effected.

(79) The dosage/effect correlation is reasonably good even if not perfectly linear.

(80) Again, it should be noted that the chestnut tannin extract, obtained according to the process described, starting from a natural raw material and without the aid of chemical substances, demonstrates its capacity of controlling attack by mites on a crop of great diffusion and importance, better than established synthesis products on the market.

(81) b) Activity on Mobile Forms of Tetranychus sp. (Expressed as % of Non-Verified Attack)

(82) The test was carried out as indicated in item a) and the results of the activity of the products tested on the mobile forms are indicated in Table 7 and in the relative graph of FIG. 9. The percentages indicated in Table 7 correspond to the percentages of crop not attacked by the mobile forms of the mites: if the crop was not attacked by the mite, the percentage is equal to 100%, whereas if the whole crop was attacked, the percentage is 0%.

(83) The evaluation was carried out on a sample of 25 leaves per block.

(84) TABLE-US-00008 TABLE 7 Effectiveness of the Formulation of Example 2 on mobile forms of Tetranychus sp. on soybeans 13-Aug. 18-Aug. 25-Aug. Days after application (DAA) (August 11) 2 7 14 Non-treated blank 0.0 0.0 0.0 Formulation of Example 2 33.2 23.9 10.2 1 Kg/ha Formulation of Example 2 39.8 39.2 16.7 2 Kg/ha Formulation of Example 2 39.5 34.4 48.6 4 kg/ha Formulation of Example 2 68.2 70.9 72.8 8 kg/ha MATACAR FL 120 g/ha 52.9 47.1 67.7

(85) It is evident from the relative graph, in FIG. 9, that the chestnut tannin extract at a dosage of 8 kg/ha is much more performing than the reference product in all the tests effected.

(86) The dosage/effect correlation is fairly good and relatively linear.

(87) This test b) also confirms what is indicated above for test a): the chestnut tannin extract demonstrates, at the highest dosage under examination, a much better control of the attack of the mite with respect to Matacar FL, whose dosage of 120 g/ha is that recommended on the label.

(88) c) Activity on the Total Mobile Forms of Tetranychus sp. (Expressed as Total Number of Mobile Forms)

(89) The test was carried out as indicated in item a) and the results of the activity on the total mobile forms are indicated in table 8 and in the relative graph of FIG. 10.

(90) The numbers indicated in Table 8 therefore correspond to the number of total mobile forms, i.e. to the number of living forms still present on the crop: the closer the value found is to zero, the more effective the treatment has been, as most of the population of mites has been eradicated.

(91) The evaluation was carried out on 25 leaves per block.

(92) TABLE-US-00009 TABLE 8 Effectiveness of the Formulation of Example 2 on the total mobile forms of Tetranychus sp. on soybeans 13-Aug. 18-Aug. 25-Aug. Days after application (DAA) (August 11) 2 7 14 Non-treated blank 563.6 544.8 468.8 Formulation of Example 2 368.8 415.7 432.8 1 Kg/ha Formulation of Example 2 381.5 349.3 422.3 2 Kg/ha Formulation of Example 2 342.6 332.6 218.3 4 kg/ha Formulation of Example 2 174.8 160.7 151.5 8 kg/ha MATACAR FL 120 g/ha 255.2 275.3 144.8

(93) It is evident from the relative graph, in FIG. 10, that the chestnut tannin extract at a dosage of 8 kg/ha is much more performing than the reference product in all the tests effected.

(94) The dosage/effect correlation is good at the highest dosages. The results indicated above for tests a) and b) are confirmed.

(95) d) Evaluation of the General Phytotoxicity Shown by the Products Tested

(96) The test was carried out as indicated in item a). The objective of this test is to evaluate and quantify any form of phytotoxicity (chlorosis, necrosis, abnormal staining, whitening, etc.) that can appear on the crop under examination following application of the products tested, at time 0 and 2 and 7 days after the treatment.

(97) The values indicated in Table 9 correspond to the percentage of plant that has suffered damage (chlorosis, necrosis, abnormal staining, whitening, etc.): the closer the value found is to zero, the more the treatment has proved to be non-phytotoxic.

(98) TABLE-US-00010 TABLE 9 11-Aug 13-Aug 18-Aug Days after application (DAA) (August 11) 0 2 7 Non-treated blank 0.0 0.0 0.0 Formulation of Example 2 0.0 0.0 0.0 1 Kg/ha Formulation of Example 2 0.0 0.0 0.0 2 Kg/ha Formulation of Example 2 0.0 0.0 0.0 4 Kg/ha Formulation of Example 2 0.0 0.0 0.0 8 Kg/ha MATACAR FL 120 g/ha 0.0 0.0 0.0

(99) It is evident that the tannin extract object of the invention, at any dosage, does not show any phytotoxicity with respect to the crop. Chlorosis, necrosis, abnormal staining, whitening, were tested.

(100) Captions

(101) FIG. 1

(102) Chromatographic profile relating to Fraction 5, registered at 280 nm: 1. Gallic acid, 2. Mono-galloyl glucose; 3. Gallotannin m/z 677; 4. Penta-galloyl glucose; 5. Galloyl-HHDP glucose; 6. HHDP glucose; 7. Ellagitannin m/z 925; 8 Castalagin/vescalagin; 9. Ellagitannin m/z 1085; 10. Ellagic acid.

(103) FIG. 2

(104) Chromatographic profile relating to the aqueous dispersion of Fraction 8. (spray dried) registered at 280 nm: 1. Mono-galloyl glucose; 2. Gallic acid 3. Digalloyl glucose; 4. Trigalloyl glucose; 5. Tetragalloyl glucose; 6. Pedunculagina isomer; 7 Ellagitannin m/z 683; 8. Ellagitannine m/z 925; 9. Castalagin/vescalagin; 10 Ellagitannine m/z 613; 11. Galloyl-HHDP glucose; 12. Ellagic acid.