WO2020229851A2

WO2020229851A2 - Edible veterinary intramammary teat sealant formulations and method for their preparation

Info

Publication number: WO2020229851A2
Application number: PCT/GR2020/000024
Authority: WO
Inventors: Georgios ARSENOS; Dimitrios FATOUROS; Eleftherios ANDRIOTIS; Evangelos KIOSIS
Original assignee: Aristotle University Of Thessaloniki - E.L.K.E.
Priority date: 2019-05-15
Filing date: 2020-05-15
Publication date: 2020-11-19
Also published as: WO2020229851A3; EP3968954A2; GR1010072B; GR20190100213A

Abstract

The present invention refers to a novel template intramammary formulation namely Mastin, as well as to a process for their preparation, wherein said formulation comprises oleogels composed of at least 75% vegetable oils and at least 5% of a food grade gelator, oleogels composed of w/o emulsions of vegetable oils, oleogels composed of suspensions of hydrophilic compounds in vegetable oils, oleogels composed of enriched vegetable oils with plant extracts, oleogels composed of vegetable oils with honey and/or its derivatives and biphasic gels composed of mixtures of vegetable oil oleogel mixed with hydrogels.

Description

EDIBLE VETERINARY INTRAMAMMARY TEAT SEALANT FORMULATIONS AND METHOD FOR THEIR PREPARATION

Field of the invention

The present invention relates to template intramammary formulations, which comprise oleogels composed of vegetable oils and of a food grade gelator, oleogels composed of w/o emulsions of vegetable oils, oleogels composed of suspensions of hydrophilic compounds in vegetable oils, oleogels composed of enriched vegetable oils with plant extracts, oleogels composed of vegetable oils with honey and/or its derivatives and biphasic gels composed of mixtures of vegetable oil oleogel mixed with hydrogels.

Background of the invention

The application of intramammary formulations is mainly focused on the treatment of various conditions associated with the mammary gland, e.g. Infections. The majority of uses of such formulations is focused on treating lactating animals. The standard method of application is infusion; the formulation is inserted into the mammary gland through the teat canal. The latter are described as injectable formulations. However, other formulations such as creams, lotions and ointments are applied topically in the skin of teats or the udder.

The most common and most serious -potentially lethal- condition of the mammary gland in dairy animals is mastitis. Mastitis is a major health problem of dairy animals worldwide. Most of the available information on mastitis relates to dairy cattle.

Some types of pathogens are set out hereafter. The inflammation of the mammary gland, whatever the origin, severity and evolution, is described as mastitis. From a clinical point of view, it can be classified as clinical or sub-clinical, depending on the extension of the microbial infection to the mammary gland. The main mastitis causing pathogens are Escherichia coli ( E . coli ), Streptococcus uberis and Staphylococcus aureus, along with a wide variety of other organisms that have been identified as potential mastitis pathogens. These organisms are described as major pathogens and are generally regarded as those commonly associated with clinical mastitis in dairy cattle.

Besides the causative microbiological factors, an important role in mastitis is also played by certain predisposing factors, such as environmental, genetics, anatomic and nutritional. Usually mastitis occurs when bacteria pass through the teat canal and enter the cistern of the mammary gland. The latter can take place either during lactation or during the dry period. During lactation, the infection can be initiated by bacteria that are present at the exterior of the teat. At the end of the milking process, the teat canal remains dilated for 1-2 hours, while damaged teats may remain open, leaving the mammary gland exposed to new infections.

The importance of mastitis is mainly economic -such as reduced milk production and treatment costs- , but also hygienic -notably with regard to dairy products-, and legal (E.U. Directives 46/92 and 71/94 defining the bacteriological quality of milk). All those issues may affect directly the dairy industry and the sustainability of dairy farms, because animals with mastitisr are prone to re-infection subject to partially recovery or permanent damage of the mammary gland tissue.

In practice, veterinarians deal with mastitis cases by administering antibiotics directly into the mammary gland using dedicated syringes. The main disadvantages of using antibiotics are the development of resistant strains of microbes, the high cost of treatment and the concerns of consumers regarding the possibility of antibiotics residues in milk if designated withdrawal periods are not implemented.

Prior art

There are a number of publications disclosing the development of intramammary teat sealant formulations for the treatment of bovine mastitis.

Both US7906138 and US 8795714 B2 describe an intramammary teat sealant and a corresponding method of forming a physical barrier in the teat canal of a non-human animal for prophylactic treatment of mammary disorders during the animal's dry period. The method includes the steps of infusing a bismuth-free teat seal formulation into the teat canal of the animal. The method also prevents the formation of black spot defect in dairy products, especially Cheddar cheese, made from the milk of animals so treated.

US6340469 describes an anti-infective free formulation for prophylactic treatment of mastitis in dry cows comprising a seal formulation having approximately 65% by weight of bismuth subnitrate in a gel based on aluminum stearate. The seal formulation is prepared by adding the bismuth subnitrate to the gel base in at least two separate stages. EP 2 369 937 B1 describes a teat sealant formulation comprising plant oil in their broadest meaning not exclusively to oil extracts, bismuth subnitrate, liquid paraffin, aluminum stearate and silicon dioxide, as a teat sealant to prevent or treat mastitis in an animal.

EP 0 971 690 B2 describes the use of a seal formulation in the preparation of a medicament for forming a physical barrier in a teat canal, particularly for the prophylactic treatment of mastitis in cows, without use of antibiotics.

US6106838 describes the preparation of various forms of pharmaceuticals for medical and veterinary uses, said pharmaceuticals comprise as active ingredients special herbal essences, capable of substituting antibiotics and sulphamide based drugs, due to their important activity against germs causing inflammations, infections and diarrhea in humans and animals. It teaches that essential oils with high content of thymol and carvacol obtained from e.g. thymus, oregano and others, are effective antibiotics for treating mastitis used in Vaseline-based treat sealants.

However, none of the above formulations can be considered edible because the raw materials used for their formulation and the final product are not food grade. Hence, all the above formulations must be removed from the animal’s teat canal prior to onset of lactation since they are not suitable for immediate consumption, e.g. by newborn animals. This is a drawback, which needs to be remedied to ensure a safe use.

Hence, one of the major concerns regarding the existing commercial intramammary formulations is the fact that they must be removed from said teat canal prior to commencement of lactation because the first milk must be discarded, as it is not safe to be consumed by young animals or even by human consumers as well. This represents additional side work which is time consuming and costs, which reduces productivity and efficiency.

Aim of the invention

It is aimed at with the present invention to bring a remedy to the abovementioned drawback. Accordingly, the present invention is an alternative proposing a timely feasible solution to this problem. The formulations that are described herein are made by food grade raw materials which are totally safe to be consumed either by young animals or by humans consuming milk or dairy products.

A product responding thereto consists of an edible product comprising bismuth subnitrate and a plant oil with anti-infective properties which is constituted by so-called Mastin. The technical aim of said Mastin is thus to provide an alternative teat sealant formulation by virtue whereof there is no need to remove it from the animal’s udder on which it is applied for treatment. This is a fundamental difference with any other existing product in the same field.

Hence, bovine mastitis is used as a model case in the context of this invention. However, the present invention is not limited to a single application, whereas it covers the whole spectrum of applications in the mammary gland.

Summary of the invention

There is thus proposed according to the invention a formulation as defined in main claim 1 as an intramammary teat sealant formulation especially for animals, particularly dairy animals, comprising at least one plant oil, at least one gelator, at least one surfactant, at least one stabilizer, at least one hydrogel and at least one w/o emulsion or suspension, which is remarkable in that the formulation is a structured plant oil or a paste or a gel or a biphasic system of an oleogel and a hydrogel forming a so- called blgel, wherein said plant oil is referred to any plant oil capable of forming oleogels; wherein said gelator is referred to any compound capable of forming oleogels; wherein said hydrogel is referred to any hydrogel known to the pharmaceutical arts.

Thanks to the invention, there is thus provided a formulation that is edible and thus safe for intramammary applications. The formulation is designed to treat or prevent inflictions of udder tissue, such as mastitis.

According to a preferred embodiment of the invention, there is proposed a veterinary or pharmaceutical formulation comprising one or more food grade plant oils and food grade gelator. The formulation is used as an intramammary infusion to prevent or treat mammary gland infections in a non-human animal.

According to an advantageous feature of said embodiment of the invention, the formulations proposed herein forms a physical barrier that inhibits the migration of bacteria from the exterior of the teat into the teat canal and finally into the cistern, thus securing the whole dairy system, from teat to cistern, to avoid therewith that it could be contaminated by said bacteria.

According to a further advantageous feature of said embodiment of the invention, the formulation provided herewith promotes wound healing and restoration of udder fatty acid balance, in the case of an infected udder. According to a still further advantageous feature of said embodiment of the invention, the formulation proposed herewith acts as antimicrobial and/or antivirus agents.

According to a yet further advantageous feature of said embodiment of the invention, the formulation proposed herewith is food grade and edible: there is thus no need for it to be removed from the teat canal prior to milking the animal and there is no concern for potential residues of harmful substances or antibiotics in milk, which thus considerably simplifies the overall milking process.

According to a particular embodiment of the invention, the formulation is based on oleogels composed of at least 75% vegetable oils and at least 5% of a food grade gelator and it is a gel or paste.

According to a preferred embodiment of the invention, the formulations contains natural food grade plant oils and food grade gelators, which makes it more sustainable.

According to a further embodiment of the invention, there is also provided a method for their manufacture and applications of the formulation proposed herein, in particular by a method of contacting or mixing the components of the formulation to form a gel or a paste or a biphasic gel.

The present invention further relates to a method for preventing or treating inflictions of udder tissue in a non-human animal. There is thus proposed according to the invention a method by which said formulation is applied to udder tissue to prevent or treat infections like mastitis.

The present invention also relates to an equipment for contacting or mixing the components of the formulation to form a gel or a paste or a biphasic gel, in short designated as bigel, including known ones.

Further advantages and features of the invention will emerge from additional subclaims appended herewith. Further details of the invention are set out in the description of some embodiments or formulations of the invention hereafter, additionally with reference to examples of preferred formulations.

Brief description of the drawings Figure 1 is a representation of extra virgin oil and beeswax oleogels with 5, 10, 15, 20 and 25 % of beeswax.

Figure 2 is a graphical representation of a textural analysis.

Figure 3 is a representation of extra virgin oil and candelillawax oleogels with 5, 10, 15, 20 and 25 % of candelilla wax.

Figure 4 shows a phase separation of extra virgin oil and candelilla wax oleogels with 5, 7,5, 10, 12,5 and 15 % of candelilla wax.

Figure 5 shows a temperature Capture at Day -1 , with a typical thermal camera image of an udder of an animal of reference.

Figure 6 shows a typical ultrasound image of the normal physiology of a teat at Day -1 of an animal of reference

Figure 7 shows a typical endoscopic image of the normal physiology of a teat cistern at Day -1 of an animal of reference.

Figure 8 shows a typical ultrasound image of a teat that is treated with a so-called "Mastin” intramammary formulation according to the invention at Day 0.

Figure 9 shows a typical ultrasound image of a teat treated with said “Mastin” intramammary formulation according to the invention after 24 h referred to as Day 1.

Figure 10 shows a typical ultrasound image of a teat treated with said "Mastin” intramammary formulation according to the invention after 48 h referred to as Day 2.

Figure 11 shows a typical ultrasound image of a teat treated with said "Mastin” intramammary formulation according to the invention after 72 h referred to as Day 3.

Figure 12 shows a typical ultrasound image of a teat treated with said "Mastin” intramammary formulation according to the invention after 96 referred to as Day 4.

Figure 13 shows a typical ultrasound image of a teat treated with said "Mastin” intramammary formulation according to the invention after 120 hours referred to as Day 5, (a) before and (b) after animal milking.

Figure 14 shows a typical endoscopic image of the normal physiology of a teat cistern that is treated with said Mastin, Day 5 after animal milking.

Description

Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The singular terms "a," "an," and "the" include plural referents unless context clearly indicates otherwise. Similarly, the word "or" is intended to include "and" unless the context clearly indicate otherwise.

The term "animal" is used herein to include all mammals, birds and fish and also include all vertebrate animals, including humans. It also includes an individual animal in all stages of development, including embryonic and fetal stages. As used herein, the term "livestock animals" refers to any breed or population of animals kept by humans as pets or for commercial purpose. For example, such animals include, but are not limited to, cattle (bovine), sheep (ovine), goat (carpa), pigs (porcine or swine), poultry (avian), and the like. As used herein, the term "cow" or "cattle" is used generally to refer to an animal of bovine origin of any age. Interchangeable terms include "bovine", "calf", "steer", "bull", "heifer”, "cow" and the like. As used herein, the term "pig" is used generally to refer to an animal of porcine origin of any age. Interchangeable terms include "piglet", "sow" and the like.

The present invention relates to veterinary or pharmaceutical formulations comprising one or more food grade plant oils and food grade gelator.

The present invention comprises food grade gelators, that form an oleogel when used in combination with a plant oil. The term oleogel or organogel includes all the gel composed of a liquid organic phase within a three-dimensional network and/or all the structured oils, i.e. oils that have been solidified in the presence of a gelator. The term gelator includes all the substances that can form an oleogel with a plant oil, such as natural and/or synthetic waxes, plant derived waxes, animal derived waxes, natural and/or synthetic polymers, nanoparticles.

All oleogels are by definition solid systems. More specifically, oleogels that are prepared by mixing vegetables oils and food grade waxes, are hard waxy solids that do not flow. Despite this fact, it was surprisingly observed herewith that when a solid oleogel is subjected to low shear mixing, the system starts to behave like a thick paste that gets thinner with every low shear mixing cycle. This remarkable behavior is attributed to the partial breakdown of the three-dimensional network of the gel that leads to a partial phase separation.

The oleogels that are made by beeswax tend to be stiffer leading to thicker and more stable pastes. The oleogels that are made by carnauba wax and/or cantdelllla wax are more prone to phase separation and the free oil that is released from the gel is used as a cleanser for the teat canal.

Additionally, the fact that the system gets thinner with every cycle of applied pressure makes the removal of formulation from the teat canal easier with every kneading of the teat.

The invention is described with reference to examples of preferred embodiments of formulations, wherein the examples below refer to the use of food grade vegetable oils, and food grade waxes of natural origin. This should not be seen as limiting as it should be appreciated by those skilled in the art that other types of gelator and/or natural ingredients may also be used in accordance with the present invention. Example 1

To assess the ability of potential food grade oils and food grade gelators to work in the required manner, a base formulation was developed. Food grade extra virgin olive oil is used as a model vegetable oil, while beeswax is used as a model food grade gelator of natural origin. This formulation was as follows:

Table 1 below shows a Base Intramammary Formulation

Table 1

The formulation was prepared according to the process described below:

1. loading virgin olive oil in a suitable vessel equipped with a low shear mixer -to avoid aeration that can promote oxidation- and starting to heat, preferably under inert atmosphere to avoid any possible oxidation reactions;

2. gradually loading the beeswax in the form of pellets and continuing to heat up to 70°C which is 5°C above the melting point of the wax, and continuing to stir until the complete dissolution of the beeswax. After the dissolution of the beeswax, the solution should be clear and transparent with no visible floating solids. Avoiding heating the mixture above 80°C to prevent discoloration of the wax;

3. stopping to stir the mixture and let the system to cool gradually. Avoiding quenching that can alter the homogeneity of the final oleogel;

4. the final oleogel is in the form of a solid waxy material that should not flow.

Oleogels with 5% and less beeswax tend to collapse under their own weight and so they are not preferred as shown in Figure 1 ;

5. the solid oleogels are then subjected to a low shear mixing process that transforms them into thick and not free flowing pastes. The mixing process may include a low shear mixing vessel with a suitable stirrer or a static mixer or any other kind of mixing equipment known to the art suitable to deliver the desirable outcome. The rheological properties of the resulted pastes are highly dependent on the total number of mixings. This dependency is depicted in Figure 2. It is clear from the textural analysis diagram that for every cycle of applied force to the oleogel, the resistance of the material to the flow is decreasing and it is mainly due to back extrusion flow of the paste around the probe, Oleogels with more than 25% beeswax are too hard to be processed and so they are not preferred;

6. 5 mL syringes are filled with the oleogel paste and they were sterilized with g radiation. The material can flow through the orifice of the syringe with the application of a pressure of maximum 10 kPa. The term syringe orifice refers to the typical syringe tips that are used for intramammary infusion applications. Example 2

This formulation was as following:

Table 2 below shows a Base Intramammary Formulation with a fraction of free oil:

The formulation was prepared according to the process described below:

1. virgin olive oil is loaded in a suitable vessel equipped with a low shear mixer -to avoid aeration that can promote oxidation- and start heating, preferably under inert atmosphere to avoid any possible oxidation reactions;

2. the candelilla wax is gradually loaded in the form of pellets and it is continued to heat up to 75°C, which is 5°C above the melting point of the wax, and stirring is continued until the complete dissolution of the candelilla wax. After the dissolution of the candelilla wax, the solution should be clear and transparent with no visible floating solids. Heating the mixture above 80°C is avoided to prevent discoloration of the wax;

3. stirring of the mixture is stopped and the system is left to cool gradually. Quenching that can alter the homogeneity of the final oleogel is avoided;

4. the final oleogel is in the form of a solid waxy material that should not flow. Oleogels with 5% and less candelilla wax tend to collapse under their own weight and so they are not preferred as represented in Figure 3;

5. the solid oleogels are then subjected to a low shear mixing process that transforms them into thick and not free flowing pastes. The mixing process may include a low shear mixing vessel with a suitable stirrer or a static mixer or any other kind of mixing equipment known to the art suitable to deliver the desirable outcome. The rheological properties of the resulted pastes are similar to those of the oleogels made with extra virgin oil and beeswax. Oleogels with more than 25% wax are too hard to be processed and so they are not preferred;

6. 5 mL syringes are filled with the oleogel paste and they were sterilized with y- radiation. The material can flow through the orifice of the syringe with the application of a pressure of maximum 10 kPa; 7. oleogels made with candelilla wax and extra virgin olive oil show a mild phase separation -free flowing oil- that is proportional to the concentration of the oil represented in Figure 3. A controlled phase separation, as in the case of 15% of candelilla wax, is desired as the free-flowing oil can serve as a cleanser that can remove any foreign object from the teat canal. Example 3

The example below refers to the use of food grade vegetable oils, and food grade waxes of natural origin, in combination with food grade hydrogels. This should not be seen as limiting as it should be appreciated by those skilled in the art that other types of vegetable oils, gelators, hydrogels and/or other natural ingredients may also be used in accordance with the present invention.

For the purpose of the present example, a formulation comprising extra virgin oil, beeswax and aloe vera hydrogel, is described. This formulation is as following:

Table 3

The formulation was prepared according to the process described below:

1. virgin olive oil is loaded in a suitable vessel equipped with a low shear mixer -to avoid aeration that can promote oxidation- and heating is started, preferably under inert atmosphere to avoid any possible oxidation reactions;

2. the beeswax is gradually loaded in the form of pellets and heating is continued up to 70°C, which is 5°C above the melting point of the wax, and stirring is continued until the complete dissolution of the beeswax. After the dissolution of the beeswax, the solution should be clear and transparent with no visible floating solids. Heating the mixture above 80°C is avoided to prevent discoloration of the wax;

4. the final oleogel is in the form of a solid waxy material that should not flow. The solid oleogel is then subjected to a low shear mixing process in order to produce a thick and not free flowing paste. The mixing process may include a low shear mixing vessel with a suitable stirrer or a static mixer or any other kind of mixing equipment known to the art suitable to deliver the desirable outcome;

5. the oleogel is stirred until a homogeneous paste is obtained with no visible defects like lumps or coagulums and then the aloe Vera gel is gradually added. Stirring until a homogeneous paste is reached with no visible islets of any of the two phases, oleogel or hydrogel;

6. 5 mL syringes are filled with the oleogel paste and they were sterilized with g radiation. The material can flow through the orifice of the syringe with the application of a pressure of maximum 7 kPa. The final biphasic gel referred to as bigel combines the properties of both gels, but for 20% concentration of the hydrogel, the paste has rheological properties which are similar to the ones of oleogel. For higher concentrations of the hydrogel, the paste starts to lose cohesion and starts to flow freely.

Example 4

The example below refers to the use of food grade vegetable oils, and food grade waxes of natural origin, in combination with food grade hydrophilic substance. This should not be seen as limiting as it should be appreciated by those skilled in the art that other types of vegetable oils, gelators and/or other natural ingredients may also be used in accordance with the present invention.

For the purpose of the present example, a formulation comprising food grade coconut oil, beeswax and pharmaceutical grade manuka honey (as the hydrophilic substance), is described. This formulation is as following:

Table 4

The formulation was prepared according to the process described below:

1. the coconut oil is loaded in a suitable vessel equipped with a low shear mixer -to avoid aeration- and heating is started, preferably under Inert atmosphere to avoid discoloration of the coconut oil;

2. the beeswax is loaded gradually in the form of pellets and heating is continued up to 70°C -5°C above the melting point of the wax- and stirring is continued until the complete dissolution of the beeswax. After the dissolution of the beeswax the solution should be clear and transparent with no visible floating solids. Heating the mixture above 80°C is avoided to prevent discoloration of the wax;

4. the final oleogel is in the form of a white solid waxy material that should not flow. The solid oleogel are then subjected to a low shear mixing process in order to produce a thick and not free flowing paste. The mixing process may include a low shear mixing vessel with a suitable stirrer or a static mixer or any other kind of mixing equipment known to the art suitable to deliver the desirable outcome;

5. the oleogel Is stirred until a homogeneous paste is achieved with no visible defects like lumps or coaguium and then the manuka honey is added gradually. Stirring is continued until a homogeneous paste is obtained with no visible islets of any of the two phases (oleogel or honey).

6. 5 mL syringes are filled with the oleogel paste and they were sterilized with g radiation. The material can flow through the orifice of the syringe with the application of a pressure of maximum 15 kPa.

The final biphasic gel has rheological properties similar to the ones of oleogel. For a higher concentration of the honey, the paste becomes sticky due to the excess amount of honey.

Example 5

The example below refers to the use of food grade w/o emulsions (or suspension of hydrophilic particles) of vegetable oils and food grade waxes of natural origin. This should not be seen as limiting as it should be appreciated by those skilled in the art that other types of vegetable oils, geiators and/or other natural ingredients may also be used in accordance with the present invention.

For the purpose of the present example, a formulation comprising food grade extra virgin oil, beeswax, pharmaceutical grade manuka honey and a non-ionic surfactant, is described. This formulation is as following:

Table 5

The formulation was prepared according to the process described below:

2. the Span 85 is added and the mixture is preheated to 45°C under continuous stirring; 3. the mixture is transferred to a vessel equipped with a high shear homogenizer. The high shear homogenization process may include any kind of homogenization equipment known to the art suitable to deliver the desirable outcome;

4. the honey is preheated to 45°C -to lower its viscosity- and it is added gradually to the extra virgin oil mixture under continuous homogenization;

5. the resulted w/o emulsion is then transferred to a suitable vessel equipped with a low shear mixer -to avoid aeration that can promote oxidation- and it is heated, preferably under inert atmosphere to avoid any possible oxidation reactions;

6. the beeswax is loaded gradually in the form of pellets and heating is continued up to 70°C -5°C above the melting point of the wax- and stirring is continued until the complete dissolution of the beeswax. After the dissolution of the beeswax, heating the mixture above 80°C is avoided to prevent creaming of the emulsion;

7. stirring of the mixture is stopped and the system is left to cool gradually. Quenching that can alter the homogeneity of the final oleogel is avoided;

8. the solid oleogels are then subjected to a low shear mixing process that transforms them into thick and not free flowing pastes. The mixing process may include a low shear mixing vessel with a suitable stirrer or a static mixer or any other kind of mixing equipment known to the art suitable to deliver the desirable outcome;

9. 5 mL syringes are filled with the oleogel paste and they were sterilized with g radiation. The material can flow through the orifice of the syringe with the application of maximum 10 kPa of pressure. The term syringe orifice refers to the typical syringe tips that are used for intramammary infusion applications.

Example 6

The example below refers to the use of enriched food grade vegetable oils with food grade plant extracts, and food grade waxes of natural origin. This should not be seen as limiting as it should be appreciated by those skilled in the art that other types of gelators and/or natural ingredients may also be used in accordance with the present invention.

This formulation is as following:

Table 6

The formulation was prepared according to the process described below: 1. virgin olive oil Hypericum Perforatum Extract is loaded in a suitable vessel equipped with a low shear mixer -to avoid aeration that can promote oxidation- and heating is started, preferably under inert atmosphere to avoid any possible oxidation reactions;

2. the candelilla wax is loaded gradually in the form of pellets and heating up is continued to 75°C (5°C above the melting point of the wax) and stirring is continued until the complete dissolution of the Candelilla wax (CW). After the dissolution of the Candelilla-wax (CW), the solution should be clear and transparent with no visible floating solids. Heating the mixture above 80°C is avoided to prevent discoloration of the wax;

4. the final oleogel is in the form of a solid waxy material that should not flow. Oleogels with 5% and less Candelilla wax (CW)tend to collapse under their own weight and so they are not preferred as represented in Figure 2;

5. the solid oleogels are then subjected to a low shear mixing process that transforms them into thick and not free flowing pastes. The mixing process may include a low shear mixing vessel with a suitable stirrer or a static mixer or any other kind of mixing equipment known to the art suitable to deliver the desirable outcome. The rheological properties of the resulted pastes are similar to those of the oleogels made with extra virgin oil and beeswax. Oleogels with more than 25% beeswax are too hard to be processed and so they are not preferred;

6. 5 mU syringes are filled with the oleogel paste and they were sterilized with g radiation. The material can flow through the orifice of the syringe with the application of a pressure of maximum 10 kPa;

7. oleogels made with candelilla wax and extra virgin olive oil Hypericum Perforatum Extract show a mild phase separation (like in Example 2) that is proportional to the concentration of the oil. A controlled phase separation, as in the case of 15% of candelilla wax, is desired as the free-flowing oil can serve as a cleanser that can remove any foreign object from the teat canal.

Example 7

Evaluation of the formulation of Example 1.

The study was performed in designated facilities of the Clinic of Farm animals of the School of Veterinary Medicine, Aristotle University of Thessaloniki, Greece (License number EL-54 BIO/exp-27). Eight (8) adult, clinically healthy purebred ewes of the Chios breed were used in the study. The average age of ewes was 18±2 months and their average milk yield was 1 ,6±0,3 L/day.

Prior to commencement of the in vivo study at day -1 , each ewe was subjected to a thorough clinical examination with emphasis on udder and teat condition based on the following protocols:

• a Clinical evaluation of the udder and the teats (observation, palpation)

• a California Mastitis Test (CMT)

• a Milk microbiological test • a Temperature capture by means of a thermal camera)

• a Teat Ultrasound

• a Teat Endoscopy

Thereafter, on the morning of Day 0 referred to as DO, the "Mastin'¹ product designating a formulation designed for intramammary administration was gently administered into the teat canal 11 of one teat 10 in each individual ewe, whereas another teat served as control reference for untreated.

The condition of every individual teat, the udder and the health status of ewes were monitored daily by clinical tests and ultrasound over a period of four (4) days.

On day one (D1) all teats were inoculated with a solution (500 cfu) containing a mixture of Coagulase- Negative Staphilococi (CNS) strains (St. xylosus, St. chromogenes, St simulans, St. hominis, St. caprae, St. lentus, St. intermedius, St. epidermidis ).

On day five (D5) the ewes were milked and they were examined by all aforementioned tests.

Assessment of the health status of ewes

As stated in the experimental design section, 24 h prior to the commencement of the actual study (Day -1), all ewes were examined for their udder and teat condition, by clinical evaluation (observation, palpation), California Mastitis Test (CMT). Table 7 shows the results of the microbiological test, whereas Table 8 shows the results of temperature measurements using a thermal camera, as well as ultrasound and Teat endoscopy. Moreover, Figures 4, 5 and 6 show images of the thermal camera, ultrasound and Teat endoscopy, respectively.

Table 7

Table 8

“Mastin” administration and monitoring of ewes

Day (0)

Following the assessment of health of individual ewes, the next day (Day 0), the said "Mastin” product which is a formulation designed for intramammary administration), was gently administered into the teat canal 11 of one teat in each individual ewe, whereas the other teat served as control (untreated). The condition of individual teats and the general health status of each individual ewe was closely monitored by clinical tests and ultrasound. The results of those measurements are presented in Table 9 and Figure 8.

Table 9

Day (1)

On day 1 , all teats were inoculated with a solution (500 cfu) containing a mixture of Coagulase- Negative Staphilococi (CNS) strains (St. xylosus, St. chromogenes, St. simulans, St. hominis, St. caprae, St. lentus, St. intermedius, St. epidermidis). The condition of individual teats and the general health status of each individual ewe was closely monitored by clinical tests and ultrasound, The results of those measurements are presented in Table 10 and Figure 9.

Table 10

Day (2)

The condition of individual teats and the general health status of each individual ewe was closely monitored by clinical tests and ultrasound. The results of those measurements are presented in Table 11 and Figure 10.

Table 11

Day (3) The condition of individual teats and the general health status of each individual ewe was closely monitored by clinical tests and ultrasound. The results of those measurements are presented in Table 12 and Figure 11.

Table 12

Day (4)

The condition of individual teats and the general health status of each individual ewe was closely monitored by clinical tests and ultrasound. The results of those measurements are presented in Table 13 and Figure 12.

Table 13

Day (5)

On Day 5 the ewes were clinically examined to assess udder and teat overall health. The procedure included clinical evaluation of the teat by observation and palpation), California Mastitis Test (CMT), microbiological test (Table 14), temperature capture by thermal camera) (Table 15), and Teat Ultrasound (Table 15, Figure 13a). The animals were then milked and teats were examined by ultrasound (Figure 13b) and endoscopy (Table 15, Figure 14).

Table 14

Table 15

Clinical Results

The novel intramammary formulation designated as "Mastin” can be easily administered at the site of application (teat).

The novel intramammary formulation referred to as "Mastin^* can be easily removed from site of application (teat).

The clinical condition of the animals is not affected upon said "Mastin" administration at the timescale of 5 days.

Teat canal mucosa is not affected by the administration of the novel intramammary formulation "Mastin” at the timescale of 5 days. Teat temperature is not affected by the administration of the novel intramammary formulation“Mastin’’ at the timescale of 5 days.

The novel intramammary formulation “Mastin’’ offers protection to the treated udder halve from intramammary infections at the tested period.

Consequently, it is concluded from the preceding that abovementioned proposed intramammary formulation has proven its efficiency in protecting the treated body parts of an animal such as udder or teat from intramammary infections.

Claims

1. An intramammary teat sealant formulation for animals having at least one teat, particularly dairy animals, comprising (a) at least one food grade plant oil and (b) at least one food grade gelator, characterized in that said formulation is made from food grade raw materials further comprising among at least (c) one surfactant, (d) at least one stabilizer, (e) at least one hydrogel and/or (f) at least one w/o emulsion or suspension, resp.

and in that the formulation is edible and sealant, wherein it is either a monophasic system consisting of a structured plant oil or a paste or a gel, or a biphasic system consisting of an oleogel and a hydrogel both forming a so-called blgel, wherein said oleogel is formed from said plant oil, respectively from a compound of said at least one gelator, wherein said hydrogel is of the pharmaceutical arts.

2. Veterinary intramammary teat sealant formulation according to claim 1 , characterised in that the formulation comprises a structured solid plant oil.

3. The intramammary teat sealant formulation according to claim 1 or 2, characterised In that the formulation is a combination of the said structured plant oil, particularly solid, and/or a paste and/or a gel and/or a biphasic system of said bigel.

4. The intramammary teat sealant formulation according to one of the claims 1 to 3, characterised in that said formulation consists of a product comprising bismuth subnitrate and a plant oil with anti- infective properties being designated as so-called Mastin.

5. A template intramammary formulation according to one of the claims 1 to 4, characterised in that the template intramammary formulation comprises oleogels composed of at least 75% vegetable oils and at least 5% of a food grade gelator consisting of a gel or paste, oleogels composed of w/o emulsions of vegetable oils; oleogels composed of suspensions of hydrophilic compounds in vegetable oils; oleogels composed of enriched vegetable oils with plant extracts; resp. oleogels composed of vegetable oils.

6, The intramammary formulation according to one of the preceding claims, characterised in that said formulation contains natural food grade plant oils and food grade gelators, thereby forming a safe veterinary or pharmaceutical formulation for intramammary applications.

7. The intramammary formulation according to claim 6, characterised in that said intramammary formulation comprises oleogels composed of vegetable oils with honey and/or its derivatives and biphasic gels composed of mixtures of vegetable oil oleogel mixed with hydrogels.

8. The formulation according to one of the preceding claims, characterised in that said formulation forms a physical barrier that Inhibits the migration of bacteria.

9. The formulation according to the preceding claim, characterised in that said formulation forms a physical barrier that inhibits the migration of bacteria from the exterior of said animal's teat to be treated into a teat canal (11) extending inwardly into the animal, into its cistern (12), thereby avoiding contamination thereof.

10. Method for producing the formulation as defined in any one of the claims 1 to 6, characterised in that said formulation is prepared by contacting or mixing the components of the formulation to form a gel or a paste or a biphasic gel referred to as bigel, resp. with an equipment consisting of a contacting or mixing device and in that said oleogel is prepared by mixing at least one vegetable oil and at least one food grade wax yielding a hard not flowing waxy solid, which is subsequently subjected to a low shear mixing, wherein said oleogel consists of an organogel including a gel composed of a liquid organic phase within a three dimensional network and at least one structured oil consisting of an oil which is solidified in the presence of a gelator consisting of a substance including a wax, natural and/or synthetic, resp. nanoparticles, forming an oleogel with a plant oil.

11. Method according to the preceding claim, characterised in that a low shear mixing cycle is carried out on this mixture, which then behaves like a thick paste getting thinner with every low shear mixing cycle that is carried out, which is thus repeated until a desired thickness of paste is reached, wherein said behavior is generated by a partial breakdown of said tridimensional network of the gel leading to a partial phase separation of said mixture;

wherein said sealing material consists of a wax, which is natural and/or synthetic, plant or animal derived wax, a natural and/or synthetic polymer; or in that sealing material is formed by nanoparticles.

12. Method according to the preceding claim, characterized in that the system formed by said oleogel is made thinner by performing successive cycles of applied pressure, after which the said formulation is removed from the teat canal (11) of the animal by exerting successive pressures while kneading the teat of the animal being treated.

13. Method for preparing the formulation according to one of the claims 10 to 12, characterized in that said formulation is prepared by the following steps:

a) loading virgin olive oil in a vessel equipped with a low shear mixer -to avoid aeration and oxidation- and starting to heat, preferably under inert atmosphere to avoid oxidation reactions;

b) gradually loading the beeswax in the form of pellets and continuing to heat up to a temperature above the melting point of the wax, in particular between 70°C and 80°C preferably, and continuing to stir this mixture until the complete dissolution of the beeswax;

c) stopping to stir the mixture and letting the whole to cool gradually, thereby avoiding quenching to enhance the homogeneity of the final oleogel, wherein

d) the final oleogel is in the form of a solid waxy consistency of not flowing material, thus forming a solid oleogel

preferably having between 5% and 25% beeswax; further wherein

e) the solid oleogels are then subjected to a low shear mixing process that transforms them into thick and not free flowing pastes, particularly wherein the mixing process includes a low shear mixing vessel with a suitable stirrer or a static mixer; further wherein

f) an injection device, esp. syringe (18) is filled with the oleogel paste, particularly wherein it (18) is sterilized with g radiation, which allows the material to flow through an orifice(19) of the syringe with the application of a limited pressure, particularly of maximum 10 kPar

14. Method according to the preceding claim, characterized in that said step (b) consists of gradually loading candelilla wax in the form of pellets and continuing to heat up to a temperature above the melting point of the wax, in particular between 75°C and 80°C preferably, and continuing to stir this mixture until the complete dissolution of the wax; particularly with about 15% of candelilla wax, yielding a controlled phase separation, wherein the free-flowing oil is released from the gel and serves as a cleanser removing any foreign object from the teat canal (11).

15. Method according to one of the claims 13 or 14, characterised in that the oleogel is stirred until a homogeneous paste is obtained with no visible defects like lumps or coagulums and then aloeVera gel is gradually added, wherein said stirring until a homogeneous paste is reached with no visible islets of any of the two phases, oleogel or hydrogel; and wherein said step (f) consists of filling an injection device, esp. syringe (18), particularly a 5 mL syringe, with the oleogel paste allowing the material to flow through its orifice (19) with the application of maximum 7 kPa of pressure.

16. Method according to one of the claims 13 or 14, characterised in that in said step (a) of the process, coconut oil is loaded in said vessel, wherein the final oleogel is in the form of a not flowing white solid waxy material, wherein the oleogel is stirred until a homogeneous paste is achieved with no visible defects like lumps or coagulum and then some honey, particularly manuka honey is added gradually, wherein stirring is continued until a homogeneous paste is obtained with no visible islets of any of the two phases oleogel or honey, wherein the oleogel paste material is allowed to flow through the orifice of the syringe with the application of a pressure of maximum 15 kPa,

17. Method according to one of the claims 13 or 14, characterised in that in said step (b) a product defined as Span 85 is added and the mixture is preheated to 45°C under continuous stirring, wherein the mixture is transferred to a vessel equipped with a high shear homogenizer, wherein the honey is preheated to 45°C to lower its viscosity and it is added gradually to the extra virgin oil mixture under continuous homogenization; wherein said syringe is filled with the oleogel paste wherein the material is allowed to flow through its orifice with the application of a pressure of maximum 10 kPa.

18. Method according to one of the claims 13 or 14, characterised in that in said step (a) virgin olive oil Hypericum Perforatum Extract is loaded in said vessel, wherein said candelilla wax is loaded gradually in the form of said pellets, wherein the material can flow through the orifice of the syringe with the application of a pressure of maximum 10kPa; wherein oleogels made with said candelilla wax and extra virgin olive oii Hypericum Perforatum Extract have a mild phase separation that is proportional to the concentration of the oil, with a controlled phase separation, particularly of 15% of candelilla wax, yielding the free-flowing oil to serve as a cleanser allowing to remove any foreign object from the teat canal (11).

19. Method for making a formulation according to one of the claims 11 to 13, characterized in that it is started from a base intramammary formulation, wherein a template model vegetable oil particularly virgin olive oil, is loaded in a container, particularly vessel, that is provided with a low shear mixing means thereby avoiding aeration and oxidation respectively, after which it is started to heat, preferably under inert atmosphere thereby avoiding oxidation reactions; after which a template model food grade gelator, particularly wax, more particularly bees wax 5 to 25%, for 95 to 75% extra virgin olive oil EVOO resp. is loaded gradually, esp. in the form of pellets, whereas heating is continued up to a temperature above the melting point of said gelator, particularly 5°C above, more particularly 70°C, in that stirring is continued until the dissolution of said gelator is complete;

after which the produced solution is made clear which is made visible by the obtained transparency thereof with no visible floating solids notably on its surface, whereas heating of the mixture is kept under 80°C maximum thereby preventing discoloration of the gelator

further wherein stirring said mixture is stopped and said system is left to cool gradually, thereby avoiding quenching to keep the homogeneity of the final oleogel;

still further wherein said final oleogel is in the form of a solid waxy not flowing material;

after which the solid oleogels are then subjected to a low shear mixing process that transforms them into thick and not free flowing pastes.

20. Method for making a formulation according to one of the preceding claims 10 to 12, in particular 14, characterized in that it is started from a base formulation, wherein a template model vegetable oil, particularly virgin olive oil is loaded in a container, particularly vessel, that is provided with a low shear mixing means thereby avoiding aeration and oxidation respectively, after which heating is started, preferably under inert atmosphere thereby avoiding oxidation reactions; after which a template model food grade gelator, particularly wax, more particularly candelllla wax, is loaded gradually, esp. in the form of pellets, whereas heating is continued up to a temperature above the melting point of said gelator, particularly 5°C above, more particularly 75⁶C,

In that stirring Is continued until the dissolution of said gelator is complete;

after which the produced solution is made clear which is made visible by the obtained transparency thereof with no visible floating solids notably on its surface, whereas heating of the mixture is kept under 80°C maximum thereby preventing discoloration of the gelator,

21. Method according to the preceding claim, characterized in that oleogel made with between 10% and 20%, preferably 15% candelllla wax and between 90% and 80%, preferably 85% extra virgin olive oil (EVOO) have a mild phase separation -free flowing oil- that is proportional to the concentration of the oil, whereas said controlled phase separation, with said 15% of candelilla wax, makes the free- flowing oil to serve as a cleanser that is brought to remove any foreign object from the teat canal (11).

22.. Method for making an intramammary formulation according to one of the preceding method claims 10 to 12, in particular 15, characterized in that it is started from a base formulation, wherein a template model food grade vegetable oil, particularly extra virgin olive oil (EVOO) and food grade wax of natural origin, esp. beeswax in combination with food grade hydrogel, esp. aloe vera hydrogel (AVG) forming a biphasic formulation for a combination of said hydrogel and oleogel, esp. composed of 64% to 80% extra virgin olive oil (EVOO), 16% to 20% beeswax and 0% to 20% AVG resp., wherein said oil is loaded in a container, particularly vessel, that is provided with a low shear mixing means thereby avoiding aeration and oxidation respectively, after which heating is started, preferably under inert atmosphere thereby avoiding oxidation reactions; after which a template model food grade gelator, particularly said wax, more particularly beeswax, is loaded gradually, esp. in the form of pellets, whereas heating is continued up to a temperature above the melting point of said gelator, particularly 5°C above, i.e. 70°C, in that stirring is continued until the dissolution of said gelator is complete;

after which the produced solution is made clear which is made visible by the obtained transparency thereof with no visible floating solids notably on its surface, whereas heating of the mixture is kept under 80°C maximum thereby preventing discoloration of the gelator, resp. said wax;

after which the solid oleogel is subjected to a low shear mixing process that transforms it into a thick and not free flowing paste;

wherein the oleogel is stirred until a homogeneous paste is obtained with no visible defects like lumps or coagulums and then the aloe Vera gel is gradually added; wherein stirring is continued until a homogeneous paste is reached with no visible islets of any of the two phases, oleogel or hydrogel; which allows the material to flow through the orifice (19) of the syringe (18) with the application of a pressure of maximum 7 kPa.

23. Method according to the preceding claim, characterized in that the final biphasic gel referred to as bigel combines the properties of both gels, said oleogel and said hydrogel (AVG), wherein the concentration of the hydrogel is chosen at 20%, by virtue whereof, the paste has rheological properties which are similar to the ones of oleogel.

24. Method according to one of the preceding method claims 10 to 12, in particular 16, characterized in that it is started from a base formulation, wherein a template model food grade vegetable oil, particularly coconut oil, and food grade wax of natural origin, esp. beeswax (BW), in combination with food grade hydrophilic substance, esp. Manuka honey (MH) hydrogel forming a biphasic intramammary formulation for a combination of honey and oleogel, esp. composed of 60% to 80% coconut oil (CO), 15% to 20% beeswax and 0% to 20% Manuka honey (MH) resp., wherein said coconut oil is loaded in a container, particularly vessel, that is provided with a low shear mixing means thereby avoiding aeration, after which heating is started, preferably under inert atmosphere thereby avoiding preventing discoloration of the coconut oil; after which said wax, particularly beeswax, is loaded gradually, esp. in the form of pellets, whereas heating is continued up to a temperature above the melting point of said wax, particularly 5°C above, i.e. 70°C,

in that stirring is continued until the dissolution of said wax is complete;

after which the produced solution is made clear which is made visible by the obtained transparency thereof with no visible floating solids notably on its surface, whereas heating of the mixture is kept under 80°C maximum thereby preventing discoloration of the gelator, resp. said wax,

further wherein stirring said mixture is stopped and said system is left to cool gradually, thereby avoiding quenching to keep the homogeneity of the final oleogel; still further wherein said final oleogel Is in the form of a solid waxy not flowing material;

wherein the oleogel is stirred until a homogeneous paste is obtained with no visible defects like lumps or coagulums and then the Manuka honey ( MH ) is gradually added; wherein stirring is continued until a homogeneous paste is reached with no visible islets of any of the two phases, oleogel or honey (MH); which allows the material to flow through the orifice (19) of the syringe (18) with the application of a pressure of maximum 15 kPa.

25. Method for making an intramammary formulation according to one of the claims 10 to 12 resp. following, characterized in that it is started from a base formulation, wherein a food grade w/o emulsion or suspension of hydrophilic particles of vegetable oil, particularly extra virgin olive oil (EVOO), and food grade wax of natural origin, esp. beeswax, in combination with pharmaceutical grade Manuka honey (MH) and a non-ionic surfactant, forming a biphasic intramammary formulation for an oleogel of a w/o emulsion, esp. composed of 60% to 80% extra virgin olive oil (EVOO), 15% to 20% beeswax 0% to 20% Manuka honey (MH) and 0% to 1 % so-called Span 85 resp., wherein said oil (EVOO) is loaded in a container, particularly vessel, that is provided with a low shear mixing means thereby avoiding aeration and oxidation respectively, after which heating is started, preferably under inert atmosphere thereby avoiding oxidation reactions; after which the said Span 85 is added and the mixture is preheated to 45°C under continuous stirring;

wherein the honey is preheated to 45°C -to lower its viscosity- and it is added gradually to the extra virgin oil mixture under continuous homogenization;

wherein the resulted w/o emulsion is then transferred to a container, particularly vessel, equipped with a low shear mixer -to avoid aeration and oxidation resp.- and it is heated, preferably under inert atmosphere to avoid oxidation reactions;

wherein the beeswax is loaded gradually, esp. in the form of pellets, and heating is continued up to 70°C - 5°C above the melting point of the wax - and stirring is continued until the complete dissolution of the beeswax; whereas after the dissolution of the beeswax, heating the mixture is kept < 80°C thus preventing creaming of the emulsion;

after which stirring of the mixture is stopped and the system is left to cool gradually, whereas the homogeneity of the final oleogel is maintained by avoiding quenching,

after which the solid oleogel is subjected to a low shear mixing process that transforms them into a thick and not free flowing paste; which allows the material to flow through the orifice (19) of the syringe (18) with the application of a pressure of maximum 10kPa.

26. Method for making a formulation according to one of the preceding claims 10 to 12, In particular 18, characterized in that it is started from a base formulation, wherein an enriched food grade vegetable oil with food grade plant extracts, particularly extra virgin olive oil Hypericum Perforatum Extract (EVOOHP), and food grade wax of natural origin, esp. candelilla wax (CW), forming a Base intramammary formulation with enriched oil, esp. composed of 75% to 95% extra virgin olive oil Hypericum Perforatum Extract (EVOOHP), and 5% to 25% candelilla wax (CW), wherein said virgin olive oil Hypericum Perforatum Extract (EVOOHP) is loaded in a container, particularly vessel, that is provided with a low shear mixing means thereby avoiding aeration and oxidation respectively, after which heating is started, preferably under inert atmosphere thereby avoiding oxidation reactions; after which said candelilla wax, is loaded gradually, esp. in the form of pellets, whereas heating is continued up to a temperature above the melting point of said wax, particularly 5°C above, more particularly 75°C,

in that stirring is continued until the dissolution of said candelilla wax is complete;

after which the produced solution is made clear which is made visible by the obtained transparency thereof with no visible floating solids notably on its surface, whereas heating of the mixture is kept under 80°C maximum thereby preventing discoloration of said wax,

after which the solid oleogel is subjected to a low shear mixing process that transforms them into a thick and not free flowing paste; which allows the material to flow through the orifice (19) of the syringe (18) with the application of a pressure of maximum 10kPa;

in that oleogel made with between 10% and 20%, preferably 15% candelilla wax and between 90% and 80%, preferably 85% extra virgin olive oil Hypericum Perforatum Extract (EVOOHP) have a mild phase separation -free flowing oil- that is proportional to the concentration of the oil, whereas said controlled phase separation, with said 15% of candelilla wax, makes the free-flowing oil to serve as a cleanser that is brought to remove any foreign object from the teat canal (11).

27. Use of the formulation as defined according to one of the claims 1 to 9 in the manufacture of a substance for the prevention or treatment of inflictions of udder tissue in a non-human animal, wherein said inflictions to be prevented consists of mastitis.

28. Use of the formulation as defined according to one of the claims 1 to 9 as an intramammary infusion to prevent or treat mammary gland infections in a non-human animal, which is applied on the animal’s organ (11 , 12, ...) to be treated by means of an injection device, notably syringe (18) being filled with the oleogel paste which is brought to flow through the orifice (19) of the syringe tip with the application of a pressure of maximum 10 kPa.

29. Use of the formulation as defined according to one of the claims 1 to 9 as antimicrobial and/or antivirus agents in a non-human animal, particularly for being left in its udder.

30. Use of said formulation in the manufacture of a substance for preventing or treating inflictions according to the preceding claim by applying said formulation to udder tissue to prevent or treat infections, resp. to promote wound healing and restoration of udder fatty acid balance, in the case of an infected udder in a non-human animal.

31. Use of the formulation as defined according to one of the claims 1 to 9 made by a method according to one of the claims 10 to 26, characterised in that said formulation is applied to udder tissue in order to prevent or treat infections consisting of mastitis.

32. Use of the free flowing oil from the gel made according to one of the claims 13 to 26, characterized in that said free flowing oil is applied to the teat canal (11) of the animal being treated as a cleanser thereof, under the action whereof any foreign and undesired object is removed from the teat canal (11)