COMPOSITION OF A SPRAY FORMULA TO CONTROL MASTITIS IN BOVINES

Abstract

The present invention corresponds to the composition of a unique formula to be used as pre-dipping and post-dipping in robot milking in order to control mastitis in bovines. The composition comprises at least one organic zinc salt, water soluble and with biocidal properties, where the concentration of zinc ions is in the range of 5,000 to 60,000 ppm. The salts with biocidal properties were selected for having a functional group similar to proteins. Organic zinc salts are degradable, less irritating to the cow's teat, improve the skin condition, have an anti-inflammatory effect, accelerate wound healing processes and eliminate bacteria and fungi. The organic zinc salts selected for this invention were the following: gluconate, lactate, glycinate, lysinate, citrate trihydrate, picolinate, and acetate. Zinc sulfate monohydrate can be optionally added, but in low concentration, less than 1.5% of the final product. The viscosity of the spray composition should be in the range of 3 to 5 centipoise (cP), at room temperature. Finally, and very importantly, the composition has no corrosive activity on metals and non-metals, such as stainless steel, carbon steel, computer cards and other components found in bovine milking robots.

Claims

1. A composition for the treatment of mastitis, wherein it comprises: a) at least one water soluble organic zinc salt selected for having a functional group similar to proteins with biocide properties at a concentration of 5,000 to 60,000 ppm of zinc ion; b) humectant agents compatible with foods; c) a viscosity agent (viscosity control agent) such that combined with humectant agents allows a target viscosity of 3 to 5 cP to be achieved; and d) demineralized water up to 100%, where the composition is non corrosive and eliminates over 99.9% of bacteria in less than 30 seconds of contact time.

2. The composition according to claim 1, wherein the water soluble organic zinc salt is selected from the group consisting of zinc gluconate, zinc glycinate, zinc lactate, zinc citrate trihydrate, zinc picolinate, zinc acetate, and zinc lysine.

3. The composition according to claim 2, wherein it preferably comprises a combination of two or more water soluble organic zinc salts.

4. The composition according to claim 3, wherein the preferred combination of zinc salts are zinc gluconate and zinc lactate in a ratio of 5/1 to 1/5 by weight, zinc gluconate and zinc glycinate in a ratio of 5/1 to 1/5 by weight, zinc gluconate and zinc acetate in a ratio of 5/1 to 1/3 by weight, zinc glycinate and zinc lactate in a ratio 3/1 to 1/3 by weight, zinc lactate and zinc acetate in a ratio of 5/1 to 1/5 by weight.

5. The composition according to claim 4, wherein the composition also comprises a third zinc salt such as zinc citrate trihydrate and/or zinc picolinate in a concentration of 1% and 0.5% by weight, respectively in the final product.

6. The composition according to claim 1, wherein the humectant agent is at a concentration between 5-20% by weight.

7. The composition according to claim 6, wherein that the humectant agents are selected from the group consisting of glycerin, propylene glycol, hexylene glycol, butylene glycol, glyceryl triacetate, aloe vera, pyrrolidone carboxylic acid, sorbitol, sodium lactate, and panthenol.

8. The composition according to claim 7, wherein the preferred humectant agent is a combination of glycerin and propylene glycol.

9. The composition according to claim 8, wherein the glycerin and propylene glycol are present in a ratio in the range of 6 to 1 to 15 to 1 by weight.

10. The composition according to claim 1, wherein the viscosity controlling agent is at a concentration range between 2-4% by weight.

11. The composition according to claim 10, wherein the preferred viscosity controlling agent is polyvinyl alcohol (PVA).

12. The composition according to claim 3, wherein one of the water soluble organic zinc salts is replaced by zinc sulfate.

13. The composition according to claim 12, wherein the preferred concentration of zinc sulfate monohydrate should be less than 1.5%, preferably less than 1.1%.

14. The composition according to claim 1, wherein it can optionally comprise additional agents selected from colorants, perfume, vitamins, alpha-hydroxy-acids, beta hydroxy-acids and hydroquinone.

15. The composition according to claim 1, wherein it is a sprayable composition for treating mastitis.

16. Use of a composition according to claim 1, wherein is used for preparing a formulation that prevents the formation of ring by hyperkeratosis of the udder tip.

17. Process for elaborating a composition for treating mastitis, wherein it comprises the following steps: a) loading demineralized water into the tank (balance at 100% formula); b) adding the soluble organic zinc salts to the tank and agitate the mixture for 10-20 minutes at 300-400 rpm until the complete dissolution of zinc salts is achieved, where all the salts are dissolved before adding the organic part in order to avoid the formation of lumps; c) adding the humectant keeping the agitation 10-20 minutes at 300-600 rpm to ensure the formation of an emulsion; d) adding the viscosity agents in the range 2% to 4% in order to achieve the target viscosity, defined as between 3 and 5 centipoise; e) agitating for 5-10 minutes at 300-600 rpm until homogeneous mixture is achieved; and f) measuring the viscosity and verifying the absence of lumps, then add coloring not exceeding a maximum concentration of 0.004%, all the stages of the process are performed at room temperature.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0036] The present invention refers to a composition to control mastitis that can be used in robot milking because it is not corrosive and has a viscosity suitable for being sprayed.

[0037] Composition

[0038] The composition is an aqueous well-balanced formula that incorporates auxiliary elements, comprising:

[0039] a) the biocide organic zinc salts, selected for having a functional group similar to proteins with biocide properties, selected from the group consisting of zinc gluconate, zinc glycinate, zinc lactate, zinc citrate trihydrate, zinc picolinate, zinc acetate, and zinc lysine; the concentration of the zinc salts being in the range of 5,000 to 60,000 ppm;

[0040] b) humectant agents suitable for foods, selected from the group consisting of glycerin, propylene glycol, hexylene glycol, butylene glycol, glyceryl triacetate, aloe vera, pyrrolidone carboxylic acid, sorbitol, sodium lactate, panthenol, among others. The preferred humectant is glycerin since it has one of the best moisture retention per weight of product ratio. In addition, glycerin has a higher viscosity (950 cP), compared with sorbitol (200 cP) and propylene glycol (52 cP), that allows for better management of the viscosity in the final product. The concentration of humectant agents in the finished product should in the range of 5-20% by weight. The most suitable humectant agents are a mixture of glycerin and propylene glycol in a weight ratio in the range of 6 to 1 to 15 to 1, as glycerin has the highest viscosity and propylene glycol creates foam, so the amount has to be restricted to the described ratio;

[0041] c) Viscosity control additives (viscosifier), such that in combination with the humectant ingredient used achieves the target viscosity in the 3-5 centipoise range, where one of the preferred additives is polyvinyl alcohol (PVA). The amount of PVA to achieve the desired viscosity is manageable (2-4% by weight), the fluid is Newtonian, does not cause phase separation and does not need to use heat or heating in the process of preparing the composition. Other thickeners with a very small amount can make the fluid of the composition (emulsion in water) become a non-Newtonian fluid with a much higher viscosity than the target, this is the case with algic acid, agar, carrageenan, pectin, gelatin, and xanthan gum. The use of these thickeners also requires a hot process to avoid lumps and achieve a stable phase over time. To reach a viscosity of 3-5 cP, these thickeners should be added in small amounts (0.1-0.3%); where solutions with these amounts are not stable over time, and the thickener tends to precipitate and separate from the solution, producing phase separation; and

[0042] d) water to form an emulsion (c.s.p. 100%)

[0043] Optionally, as previously mentioned, the suitable inorganic salt that can be added in combination with at least one organic zinc salt is zinc sulfate monohydrate in a concentration below 1.5%, preferably below 1.1% by weight.

[0044] Optionally, other agents could be added, such as colorants, perfume, vitamins, alpha-hydroxy-acids, beta hydroxy-acids, hydroquinone and other agents which aid skin treatment.

[0045] Process

[0046] The dipping solution (composition) is prepared in three stages, in the first stage the zinc salts are dissolved in water. In the second stage, the organic material (humectants and thickening agents) is added. Finally, the third stage involves the addition of the colorant.

[0047] The composition is prepared in a mix tank (a tank with an axial agitator).

[0048] The main steps of the process are: [0049] a) A mix tank is loaded with demineralised water, depending on the composition of the formula (c.s.p. 100%). [0050] b) Zinc salts are added in the range from 6% to 20% dependent upon the selected group of salts and zinc ion concentration target and the system is agitated until complete dissolution of zinc salts. The agitation time goes from 10-20 minutes at an agitation speed of 300-400 rpm until complete dissolution is achieved. [0051] c) Humectants are added in the range of 5-20%. The system is agitated until the formation of the emulsion is ensured, the agitation time is 10-20 minutes and the agitation speed is 300-600 rpm. [0052] d) Thickening agent is added in the range of 2 to 4% to achieve the target viscosity. [0053] e) Agitation is continued until a homogeneous mixture is obtained. [0054] f) A sample is taken from the tank and visually inspected with respect to the non-presence of lumps and the viscosity is measured. [0055] g) Finally, if everything fulfills the requirements a colorant solution is added: The colorant is previously dissolved in a small amount of water.

[0056] It is necessary to confirm that all zinc salts are completely dissolved before adding the humectants and thickening agents. If not, lumps may form. [0057] The order of addition is critical to obtain a dipping with the required characteristics.

[0058] The selected humectant agents are glycerin and propylene glycol, where glycerin has the best water retention among all known humectants, while propylene glycol is selected because it reduces the surface tension of dipping and acts as a stabilizer.

[0059] The selected viscosifying agent is PVA. Its concentration is between 2% and 4%, preferably in the range of 3% to 4% to obtain a viscosity of 3 to 5 cP. No other thickening agent is desirable for the production of this product: such as algic acid, agar, carrageenan, pectin, gelatin, xanthan gum, and others.

[0060] The process is conducted at room temperature, which differs with most process that use a temperature of 60° C.-80° C. It has been found that an increase in the rate of agitation over 1000 rpm promotes the polymerization of PVA, causing lumps in finished product, which reduces the shelf life of the dipping.

[0061] Biocide Organic Zinc Salts [0062] The active ingredient corresponds to organic zinc salts, where two or more salts have a higher biocidal activity than a single zinc salt in the composition. The total concentration of zinc ions resulting from the contribution of all salts should be in the range of 5,000 to 60,000 ppm, preferably in the range of 10,000 to 25,000 ppm.

[0063] Therefore, the type, amount and solubility of the selected zinc salts are relevant for the present invention in order to achieve the target zinc ion concentration with the selected salt or pairs of salts.

[0064] The selected zinc organic salts for this present innovation are: zinc gluconate, zinc glycinate, zinc lactate, zinc citrate trihydrate, zinc picolinate, zinc acetate, and zinc lysine.

[0065] The preferred mixtures of zinc salts are zinc gluconate and zinc lactate in a ratio of 5/1 to 1/5 by weight, zinc gluconate and zinc glycinate in a ratio of 5/1 to 1/5 by weight, zinc gluconate and zinc acetate in a ratio of 5/1 to 1/3 by weight. In addition the mixtures of zinc glycinate and zinc lactate in a ratio 3/1 to 1/3 by weight, zinc lactate and zinc acetate in a ratio of 5/1 to 1/5 by weight can be also used. In all these mixtures, a third zinc salt such as zinc citrate trihydrate and zinc picolinate could be added in a concentration of 1% and 0.5% by weight, respectively in the final product.

[0066] The use of 2 or more zinc salts in the mixture has proven to increase the antimicrobial performance. In addition, it provides flexibility to achieve the target zinc ion concentration in finished product given the water solubility of these salts.

[0067] Zinc Sulfate

[0068] Optionally, zinc sulfate mono-hydrate can be added in combination with at least one organic zinc salt in order to achieve a proper concentration of zinc ions. Zinc sulfate mono-hydrate is added in such a way that its concentration should be below 1.5%. The reasons for this restriction is that concentrations above that value tend to react with PVA, forming lumps. The interaction is due to the reaction within the sulfate from the zinc sulfate with the PVA.

[0069] The salt mixtures that are preferred to be mixed with zinc sulfate mono hydrate are zinc gluconate and zinc lactate; zinc gluconate and zinc glycinate; zinc gluconate and zinc acetate; zinc glycinate and zinc lactate; zinc lactate and zinc acetate. In addition, zinc citrate trihydrate and zinc picolinate could be added to the mixture, as previously indicated. In all the cases, zinc sulfate mono hydrate could be added at a concentration below 1.5% in the finished product.

[0070] Viscosity

[0071] As mentioned, a high viscosity dipping (over 10 cP) is not desirable for spraying (carousel-type milking processes). Whereas if the viscosity is close to 1 cP, the sprayed product forms a ring and does not completely wet the udders when sprayed.

[0072] Therefore, to fully wet the udders and obtain good atomization, the viscosity of the dipping should be in the range of 3 to 5 centipoise. To achieve the proper viscosity, the humectant agent can be combined with an appropriate amount of thickening agent. The viscosity of the product is achieved through the correct mixture of glycerin, propylene glycol and PVA, by forming an emulsion with the inorganic phase that corresponds mainly to water, the aqueous solution of zinc salts.

[0073] The use of the combination of glycerin and PVA responds to the fact that, although glycerin contributes to the viscosity, it was found that increasing the content in the dipping to 20% did not achieve the target viscosity Similar tests with just PVA also did not show good results, as very small or very large amounts formed lumps in the dipping and the stability of the product was lost. However, by adjusting a mixture of glycerin and PVA with a weight ratio of 1.5 to 5, it is possible to obtain a stable formula, which meets the target viscosity, and exhibits good adherence to the cow's udders.

[0074] As indicated, the spray system of the robot limits the viscosity range. A product with a viscosity greater than 10 cP cannot be sprayed. In the range of 5-10 cP only a straight stream comes out, while in the range of 3-5 cP a wide spray is obtained that moistens the entire surface of the udders. Viscosity values under 3 cP, result in a concentric circle type irrigation that does not completely wet the udders, wetting the front of the udder and not the teats.

[0075] The spray system of the robot is programmed to operate for a period of 4-6 seconds in pre-dipping and post-dipping mode. This time is sufficient to spray 15-25 ml of the dipping per cow. Therefore, the challenge for a good dipping performance is that it must work at the robot's operating conditions and must be effective in controlling mastitis through a spray that soaks all the cow's udders, which is able to adhere and seal them, and that eliminates the bacteria present in the udders.

[0076] Biocide Effectiveness

[0077] The dipping having 10,000 ppm of zinc ions, from the mixture of zinc gluconate and zinc lactate (equivalent to 3.485% zinc gluconate and 1.865% zinc lactate) was subjected to quantitative germicidal rate tests (Time Kill Kinetics Assay, ASTM E2315). Briefly, 10 ml of the dipping were inoculated with a concentrated solution of bacteria in order to reach 10.sup.6 CFU/ml, then the mixture was agitated for 30 seconds, an aliquot was taken which was seeded by flooding on Plate Count agar, to then be incubated for 24 hours at 36° C. After this, the bacterial colonies were counted and the percentage of elimination was determined with respect to a control sample where the dipping was replaced by water, maintaining the test conditions described above.

[0078] The percentage of elimination of bacteria at 30 seconds contact time was: Escherichia coli (100%), Staphylococcus aureus (100%), Streptococcus uberis (99.99%), Streptococcus agalactiae (99.98%), Streptococcus dysgalactiae (99.90%), Staphylococcus coagulasa negative (100%), and Pseudomonas spp (99.99%).

[0079] For the qualitative evaluation of the bactericidal activity of the different dipping (salt mixtures) the Kirby Bauer method (Halo Method) was used, where a plate with Mueller Hinton agar was inoculated with a bacterial concentration of 1.5×10.sup.8 CFU/ml, to then, by punching out 4 holes of 5 mm diameter per 90 mm diameter plate. Each hole was independently inoculated with 35 microliters of dipping, and then the plates were incubated for 18-20 hours at 36° C. Finally, the diameter of the halo is measured. The larger the diameter, the greater the antibacterial activity.

[0080] Non-Corrosiveness

[0081] The fact of having a non-corrosive formulation allows the present invention to be a versatile composition, which can be used in milking robots, not only as a pre-dipping but also as a post-dipping, without changing the concentration of biocide organic zinc salts nor the viscosity or any other property of the formulation. The versatility of the invention also allows for use as pre- or post-dipping in other milking systems, such as the carousel type.

[0082] It is desirable to incorporate colorant or dye in the dipping solution. This way, it is easy to detect if the post dipping was applied or not and the quality of application. We have selected the mixture of blue and green food grade colorant. The concentration of colorant in the dipping is ideally 0.001%-0.003% for blue and 0.001%-0.003% for green. Not exceeding 0.004% in total. We have found that greater concentrations of colorant stain the visor of the camera of the robot, increasing the difficulty of locating the udder of the cow.

[0083] The amount of colorant in the present invention is almost 10 times less than those described in other dipping patents.

EXAMPLES

[0084] The following examples are meant to illustrate the invention, but in no case to limit it.

Example 1

[0085] One Kilogram of product (dipping) with a composition according to the present invention is made dissolving 8.362% of zinc gluconate in 80.838% of water, while mixing at 300 rpm at room temperature. After dissolution is completed, 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA is added, and mixed for 10 minutes at 500 rpm.

[0086] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0087] The viscosity of the dipping is 3.2 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

Example 2

[0088] One Kilogram of product (dipping) with a composition according to the present invention is made dissolving 4.471% of zinc lactate in 84.729% of water, while mixing at 300 rpm at room temperature. After dissolution is completed, 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA is added, and mixed for 10 minutes at 500 rpm.

[0089] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0090] The viscosity of the dipping is 3.1 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

Example 3

[0091] One Kilogram of product (dipping) with a composition according to the present invention is made dissolving 3.368% of zinc acetate in 85.832% of water, while mixing at 300 rpm at room temperature. After dissolution is completed 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA is added, and mixed for 10 minutes at 500 rpm.

[0092] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0093] The viscosity of the dipping is 3.2 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

[0094] The diameter of the halos was measured for the following bacteria:

TABLE-US-00001 TABLE 1 Diameter of Diffusion Inhibition Halo (mm) for 12.000 ppm of zinc ions, using different zinc salts Example 1 Example 2 Example 3 Zinc salt Zinc Gluconate Zinc Lactate Zinc Acetate ppm Zinc 12.000 12.000 12.000 Staphylococcus aureus methicillin sensitive 20 18 19 ATCC 25923 Escherichia coli ATCC 25922 16 17 16 Staphylococcus aureus - Isolated from 20 20 20 Clinical Mastitis Streptococcus agalactiae - Isolated from 17 17 16 Clinical Mastitis Escherichia coli (coliforme) - Isolated from 16 16 16 Subclinical Mastitis Streptococcus uberis - Isolated from 18 17 18 Subclinical Mastitis Streptococcus agalactiae - Isolated from 18 18 17 Subclinical Mastitis

[0095] The following examples (4 to 7) use a mixture of two zinc salts. Each zinc salt contributes 6,000 ppm of zinc ion.

Example 4

[0096] One Kilogram of product (dipping) with a composition according to the present invention is obtained dissolving 4.181% of zinc gluconate, 2.235% of zinc lactate in 82.783% of water, while mixing at 300 rpm at room temperature. After dissolution is completed 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA is added, and mixed for 10 minutes at 500 rpm.

[0097] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0098] The viscosity of the dipping is 3.2 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

Example 5

[0099] One Kilogram of product (dipping) with a composition according to the present invention is obtained dissolving 4.181% of zinc gluconate, 1.684% of zinc acetate in 83.335% of water, while mixing at 300 rpm at room temperature. After dissolution is completed 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA is added, and mixed for 10 minutes at 500 rpm.

[0100] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0101] The viscosity of the dipping is 3.2 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

Example 6

[0102] One Kilogram of product (dipping) with a composition according to the present invention is obtained dissolving 2.235% of zinc lactate, 1.684% of zinc acetate in 85.281% of water, while mixing at 300 rpm at room temperature. After dissolution is completed 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA, is added, and mixed for 10 minutes at 500 rpm.

[0103] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0104] The viscosity of the dipping is 3.2 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

Example 7

[0105] One Kilogram of product (dipping) with a composition according to the present invention is obtained dissolving 1.967% of zinc glycinate, 3.371% of zinc lysinate in 83.861% of water, while mixing at 300 rpm at room temperature. After dissolution is completed 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA is added, and mixed for 10 minutes at 500 rpm.

[0106] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0107] The viscosity of the dipping is 3.2 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

TABLE-US-00002 TABLE 2 Diameter of Diffusion Inhibition Halo (mm) obtained with 12.000 ppm of zinc ions from a mixture of two zinc salts Example 4 Example 5 Example 6 Example 7 Zinc salt Zinc Zinc Zinc Lactate Zinc Glycinate Gluconate Gluconate Zinc Zinc Lysinate Zinc Lactate Zinc Acetate Glycinate ppm Zinc 12.000 12.000 12.000 12.000 Staphylococcus aureus methicillin 23 21 22 23 sensitive ATCC 25923 Escherichia coli ATCC 25922 19 20 19 21 Staphylococcus aureus - Isolated from 25 23 24 24 Clinical Mastitis Streptococcus agalactiae - Isolated 20 21 20 20 from Clinical Mastitis Escherichia coli (coliforme) - Isolated 20 19 21 20 from Subclinical Mastitis Streptococcus uberis - Isolated from 21 20 21 21 Subclinical Mastitis Streptococcus agalactiae - Isolated 21 21 20 21 from Subclinical Mastitis

[0108] Examples 8 to 10 show the results of Halo Inhibition measured for a mixture of two organic zinc salts, to which 1.098% of zinc sulfate monohydrate (a contribution of 4,000 ppm zinc ion) has been added. The total zinc ion concentration in the test is 28,000 ppm. The difference in zinc ion concentration was contributed in equal parts by the other salts, 12,000 ppm of zinc ion, each.

Example 8

[0109] One Kilogram of product (dipping) with a composition according to the present invention is obtained dissolving 8.362% of zinc gluconate, 4.471% of zinc lactate, 1.098% zinc sulfate monohydrate in 69.069% of water, while mixing at 300 rpm at room temperature. After dissolution is completed 12% of glycerin, 1% propylene glycol and 4% of PVA it is added, and mixed for 10 minutes at 500 rpm.

[0110] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0111] The viscosity of the dipping is 3.9 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

Example 9

[0112] One Kilogram of product (dipping) with a composition according to the present invention is obtained dissolving 4.471% of zinc lactate, 3.368% of zinc acetate, 1.098% zinc sulfate monohydrate in 74.063% of water, while mixing at 300 rpm at room temperature. After dissolution is completed 12% of glycerin, 1% propylene glycol and 4% of PVA is added, and mixed for 10 minutes at 500 rpm.

[0113] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0114] The viscosity of the dipping is 3.8 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

Example 10

[0115] One Kilogram of product (dipping) with a composition according to the present invention is obtained dissolving 3.934% of zinc glycinate, 6.742% of zinc lysinate, 1.098% zinc sulfate monohydrate in 71.226% of water, while mixing at 300 rpm at room temperature. After dissolution is completed 12% of glycerin, 1% propylene glycol and 4.0% of PVA is added, and mixed for 10 minutes at 500 rpm.

[0116] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0117] The viscosity of the dipping is 3.9 cP, measured with a Digital Viscometer, Model NDJ-1S at 25° C.

TABLE-US-00003 TABLE 3 Inhibition Halo Diameter (mm) obtained with 28,000 ppm of zinc ions from a mixture of two organic zinc salts plus zinc sulfate monohydrate. Example 8 Example 9 Example 10 Zinc salt Zinc Gluconate Zinc Lactate Zinc Zinc Glycinate Zinc Zinc Lactate Acetate Lysinate Zinc Zinc Sulfate Zinc Sulfate Sulfate Monohydrate Monohydrate Monohydrate ppm Zinc 28.000 28.000 28.000 Staphylococcus aureus 29 28 29 methicillin sensitive ATCC 25923 Escherichia coli ATCC 25922 26 27 26 Staphylococcus aureus - Isolated 32 31 32 from Clinical Mastitis Streptococcus agalactiae - 26 26 27 Isolated from Clinical Mastitis Escherichia coli (coliforme) - 25 26 26 Isolated from Subclinical Mastitis Streptococcus uberis - Isolated 27 26 27 from Subclinical Mastitis Streptococcus agalactiae - 26 26 26 Isolated from Subclinical Mastitis

[0118] Corrosive Effect

[0119] The corrosive effect of dipping was determined in different metallic and non-metallic parts, such as: iron, zinc-plated iron, 316 stainless steel and computer boards. Each of the pieces was placed independently in a 1 liter glass beaker containing 800 ml of dipping. The vessels were kept at 25° C. for six months, each piece being inspected visually and microscopically every two weeks for the first two months and once a month for the next 4 months. Inspections look for signs of attack on the surface, such as pitting, cracks, crevices, or color changes No signs of corrosion were observed on the parts. The dipping used is that of the examples.

[0120] Skin

[0121] An experimental test was designed to evaluate the quality of the skin and the tip of the udders, comparing them after the application of different pre and post dipping according to the following:

[0122] (a) Pre and post dipping according to the present invention (20,000 ppm of zinc ion from equal parts of zinc gluconate and zinc lactate); versus

[0123] (b) Pre and post dipping from a commercial formula based on 3% glycolic acid; and

[0124] (c) Pre and post dipping of a commercial formula based on 0.5% hydrogen peroxide and 1.7% lactic acid.

[0125] The test lasted 1 month and the evaluation panel included 100 cows per dipping, where each cow was milked three times a day, evaluating the skin quality of the 4 udders daily, marking a score for the skin of the teat and the tip of the teat according to the scale used in the standard evaluations of the industry.

TABLE-US-00004 TABLE 4 Udder Skin Quality Score after pre and post dipping application: Summary of Results Condition of Zinc Salts Glycolic acid Hydrogen peroxide + lactic acid the skin (a) (b) (c) Normal 100% 60% 20% Dry  0% 20% 25% Cracked  0% 10%  5% Irritated  0% 10% 25% Flaked  0%  0% 25%

[0126] The score was assigned according to the standard evaluation in the industry, Teat End conditions (QCW-7: Teat End Conditions Scorecard).

TABLE-US-00005 TABLE 5 Evaluation of the tip of the teat after the application of different pre- and post dipping: Summary of Results. Hydrogen peroxide + lactic acid Condition of the Tip Zinc Salts (a) Glycolic acid (b) (c) No ring 90% 50% 30% Soft ring 10% 35% 40% Wrinkled ring  0% 10% 15% Very Wrinkled ring  0%  5% 15%

[0127] The ring is formed by hyperkeratosis of the tip of the teat. The absence of a ring indicates a healthy condition.

[0128] Results show that zinc salts provide greater protection for teat tips and maintain udders in better conditions than commercial formulas based on different biocidal agents.

[0129] Examples 11-14: Comparison of the antibacterial activity of the invention versus commercial formulas based on different biocidal agents.

[0130] The antibacterial activity of the dipping composition according to the present invention was evaluated and compared with other commercial formulas based on different biocidal agents, in the same matrix solution. Antibacterial activity tests compare the diameters of the inhibition halos.

Example 11

[0131] One Kilogram of product (dipping) is obtained dissolving 2.787% of zinc gluconate, 1.49% of zinc lactate, 1.098% of zinc sulfate monohydrate. Each zinc salt contributes with 4,000 ppm of zinc to the solution for a total of 12,000 ppm zinc. An 83.825% of water, while mixing at 300 rpm at room temperature is added. After total dissolution is achieved, 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA are added and the composition is mixed for 10 minutes at 500 rpm.

[0132] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

Example 12

[0133] One Kilogram of product (dipping) is obtained dissolving 1.922% of zinc sulfate monohydrate (a total of 7,000 ppm of zinc ion) and copper sulfate pentahydrate 2.75% (a total of 7,000 ppm of copper ion). An 84.528% of water, while mixing at 300 rpm at room temperature is added. After total dissolution is achieved, 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA are added and the composition is mixed for 10 minutes at 500 rpm.

[0134] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×10.sup.8 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

Example 13

[0135] One Kilogram of product (dipping) is obtained dissolving 3% glycolic acid (a total of 30,000 ppm of glycolic acid) and 86.2% of water, at 300 rpm at room temperature. After the mixing is completed, 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA are added and the composition is mixed for 10 minutes at 500 rpm.

[0136] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×108 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

Example 14

[0137] One Kilogram of product (dipping) is obtained mixing 0.5% hydrogen peroxide, 1.7% lactic acid, and 87% of water, at 300 rpm at room temperature. After the mixing is completed, 6.5% of glycerin, 0.8% propylene glycol and 3.5% of PVA are added and the composition is mixed for 10 minutes at 500 rpm.

[0138] A volume of 35 microliters is added to the 5 mm diameter hole previously made in a Mueller Hinton plate inoculated with bacteria at a concentration of 1.5×108 UFC/ml. The diameter of diffusion inhibition halo is measured after 20 hours incubation at 36° C.

[0139] The results of the diameters of the inhibition halos (mm) are shown in the following table.

TABLE-US-00006 TABLE 6 Diameters of the Inhibition Halos (mm) obtained from the invention versus formulas based on the use of different biocidal actives Example 11 Zinc Example 14 Gluconate Example 12 Hydrogen Zinc Lactate Copper Sulfate Example 13 peroxide + Zinc Sulfate Zinc Sulfate Glycolic Acid lactic acid Staphylococcus aureus methicillin 23 8 10 20 sensitive ATCC 25923 Escherichia coli ATCC 25922 19 10 10 16 Staphylococcus aureus - Isolated from 25 10 11 20 Clinical Mastitis Streptococcus agalactiae - Isolated 20 8 12 18 from Clinical Mastitis Escherichia coli (coliforme) - Isolated 20 9 10 16 from Subclinical Mastitis Streptococcus uberis - Isolated from 21 11 12 18 Subclinical Mastitis Streptococcus agalactiae - Isolated 21 9 12 19 from Subclinical Mastitis

[0140] The results show that this invention, having 12,000 ppm of zinc ions (mixture of three zinc salts) has a higher antibacterial activity than (a) a mixture of 7,000 ppm of zinc ion plus 7,000 ppm of copper ion; (b) 3% glycolic acid; and (c) a mixture of 0.5% hydrogen peroxide and 1.7% lactic acid.

Example 15

[0141] The dipping (solution) is prepared in an agitated tank equipped with an axial agitator with variable speed up to 700 rpm and which consists of the following steps: [0142] a) A 1.5 cubic meter tank is loaded to prepare 1,000 kilograms of the present invention, adding between 770-810 kilograms of demineralized water. [0143] b) Zinc salts are added and the system is agitated from 15 minutes at 400 rpm. [0144] c) Glycerin and propylene glycol are added in a ratio from 1.5-5 by weight and with agitation for 15 minutes at 600 rpm. [0145] d) PVA is added in the range 2% to 4%. [0146] e) Agitation is continued 5 minutes at 600 rpm. [0147] f) Viscosity is measured and the absence of lumps is visually inspected. [0148] g) An aqueous solution of dye previously dissolved in water is added so that the concentration of the blue pigment is in the order of 0.001%-0.003% and of the green pigment in the same range. The sum of the pigments must not exceed 0.004%.