The invention generally relates to the field of veterinary herbal products. More specifically, it relates to a topical barrier film-forming herbal composition for the prevention and management of mastitis and method thereof.
BACKGROUND OF THE INVENTION
Mastitis is the inflammation of the mammary gland and udder tissue and is a major endemic disease resulting in huge economic loss to the dairy industry. It usually occurs as an immune response to bacterial invasion of the teat canal by variety of bacterial species such as Staphylococcus aureus, Streptococcus faecalis, Streptococcus pyogenes, Klebsiella spp., Mycobacterium bovis and E. coli (Reference :https://dairy.ahdb.org.uk/technical-information/animal-healthwelfare/mastitis). The bacterial toxins released in the udder tissue damage the milk-secreting tissues and ducts throughout the udder. In some situations, the damage is permanent and the udder loses its functionality. Acute cases can lead to fatalities while cows that recover may be lactationally incapacitated for the rest of their lives.
Mastitis in cows can be a very complex problem since the farmer may not be able to detect it during its subclinical phase (a phase of mastitis without showing any signs and symptoms). At this stage, the disease can spread very fast to the rest of the herd if proper handling is lacking in the farm. The losses a farmer/ farm-owner is likely to incur arise from the following avenues:
• Milk thrown away due to contamination by medication or being unfit to drink.
• A reduction in yield due to illness and any permanent damage to udder tissue.
• The extra labor required to tend mastitic cows.
• The costs of veterinary care and medicines.
• The cost of reduced longevity due to premature culling.
Economic loss- Globally, mastitis accounts for about 38 per cent of the total direct cost of animal farming. In India, the economic losses due to mastitis have increased about 115-130 folds in last few years. Lack of awareness, delay in detection of mastitis, lack of markers for detecting ensuing mastitis, unhygienic milking practices, diverse production systems, inadequate treatment etc. are some of the important factors contributing to higher incidence of mastitis.

Existing methods for the prevention of mastitis and their limitations
The most common method for prevention of mastitis involves the use of antiseptic or antimicrobial agents, which are chemical in nature. These include iodophors, quaternary ammonium compounds, chlorhexidine salts, chlorine-release compounds (e.g. alkali hypochlorites), oxidizing compounds (e.g. hydrogen peroxide, peracids), protonated carboxylic acids (e.g. heptanoic, octanoic, nonanoic, decanoic, undecanoic acids), acid anionics (e.g. alkyl-aryl sulfonic acids), and chlorine dioxide (from chlorite). These agents have varying degrees of effectiveness and limit the transmission of mastitis by reducing pathogen populations on the teat surface.
Method of use- by dipping teats in solution of the compounds or spraying the solution on the teats. The solutions form a layer on the outer surface of the teats. The teat skin and muscles being tough, bacteria cannot enter through skin but only through the teat canal. Limitations
i. Not efficient-due to insufficient amount of active agents or the insufficient contact time
with the teat skin leading to incomplete prevention of mastitis, ii. Recurrence of pathogens: It is also known that the effect of existing agents fades out very quickly and that renewed contamination of the teats takes place shortly after the treatment. Small wounds and skin tissues on the teats can act as reservoirs for certain microbes, and livestock with infected teat wounds often show higher mastitis frequencies than other livestock, iii. Chemicals can enter into milk: Dipping of the teats into chemical solutions e.g. iodine, chlorhexidine and other agents can result in movement of these chemicals into milk which can not only be harmful for health but also interfere with the manufacture of dairy products.
In view of the foregoing, there exists a need for compositions which are safer and not harmful for health of animal or persons consuming milk from treated animals.

Introduction to present invention
The present invention discloses a topical barrier film-forming herbal composition for the prevention and management of mastitis. The said herbal composition consists of a synergistic combination of the following viz.
i. Cedrus deodara oil- 1-3%, optimally 2%
ii. Eucalyptus globulus oil- 0.5-1.5%, optimally 1% iii. Curcuma longa oil - 0.1-0.3%, optimally 0.2% iv. Gaultheria jragrantissima oil - 0.5- 1.5%, optimally 1%
v. A natural polysaccharide, preferably tamarind seed polysaccharide - 10-14%,
optimally 12% (w/v). Mechanism of action
When the said liquid composition is applied topically, it soon dries, forming a plug at the teat orifice, which acts like a mechanical barrier, thus preventing the entry of bacteria in the udder through the teat canal. Film formation also allows better surface retention of the active components on the surface of the teats. Further the synergistic mixture of oils results in broad-spectrum antimicrobial activity that kills nearly 99.95%) of the common mastitis-causing bacteria. Further, the essential oils exert emollient (skin softening) and vulnerary (wound healing) activities on the teat skin. Advantages
i. Safety- uses natural herbal ingredients and is thus free from the undesirable effects on human health associated with chemical compounds, which when applied on the teats, contaminate milk. The composition consists of ingredients that are classified under GRAS (Generally Regarded As Safe) category and, thus, eliminate the risks of contamination of milk. II. Unique mechanism of action- The herbal composition does not remain in 'wet' form on the teat after application, unlike the chemical compound solutions. It dries quickly, forming a protective barrier film, which prevents the entry of pathogens into the teat canal. Film formation allows better surface retention of the active components on the surface of the teats. The composition kills nearly all the common microbes, which may cause mastitis, due to broad spectrum bactericidal activity of the oils.

iii. Additional benefits - the composition not only has a preventive/killing effect on microbes but also a healing effect on skin wounds and fissures, thereby strengthening and enhancing the protection.
iv. Easy application- Furthermore, the composition can be given such consistency that it is easy to apply to the teats, for example by dipping or spraying, at the same time as it imparts an emollient effect on the teats. Also, the composition is not harsh on the milkers hand or milking machine and can be peeled off the teats or washed off easily with water before milking.
v. Economical production- All the ingredients viz. 4 essential oils and tamarind seed polysaccharide are easily available commercially and cheaply which makes production of the composition economical. Prior art and limitations

S.No. Patent Application No. Prior Art Patent Present Invention
1. EP 0917427 Discloses a teat dip composition comprising of chitosan and a group of polysaccharides. The said composition could treat only 91% of the infected animals Consists of a synergistic combination of essential oils viz. Cedrus deodara oil, Eucalyptus globulus oil, Curcuma longa oil and Gaultheria jragrantissima oil and one polysaccharide, preferably tamarind seed polysaccharide in an optimum concentration. The composition of the present invention can prevent/ kill 99.95% of the common mastitis-causing microbes.
2. EP0904693 Bl Discloses a composition for treatment of mastitis which consists of an acidulant and Composition is gentle on the teat and free from any strong corrosive compounds. Rather it

chlorite. The said composition being chemical in nature can be harsh to the teats and hands of milker and can contaminate the milk. contains natural oils, which apart from antimicrobial effect, offer emollient and wound healing activities.
3. US 4945110 Discloses an antibacterial chemical composition for mastitis prevention which is an aqueous solution of an aliphatic sulfate or sulfonate salt detergent, lactic acid or a food grade salt. No chemical compound is present at all in composition of present invention.
4. US 5776479 Discloses a germicidal aqueous teat-dip composition which comprises a film-forming agent selected from the group consisting of hydroxyethylcellulose, methyl hydroxyl-propylcellulose, and ethylhydroxyethylcellulose, iodine complexed with a non-ionic surfactant and water. Composition of present invention is free from any synthetic film forming agents or chemical agents including non-ionic surfactants.
5. US 7078062 B2 Discloses an aqueous solution of hop-based udder and teat dips to wash and sanitize the teats. Composition is entirely different consisting of a synergistic mixture of 4 essential oils and a natural polysaccharide for film formation on drying.
6

6. US 2011/0086117 Al Discloses an aqueous antiseptic composition comprising only one essential oil i.e. 10 to 50% by volume of lemongrass oil and one solubilizing agent for solubilizing the lemongrass oil, 10 to 30% by volume of a carrier oil carrying the components of the composition and about 0.5 to 2% by volume of an electrolyte for providing conductivity for the com¬position. The composition further comprises an organic acid for adjusting the pH level of the composition and an antioxidant. The composition is not aqueous but oil-in-water emulsion.
7. Waghmare et al., 2013. Int. J. Agric.Sc & Vet.Med. 41-50. Discloses an aqueous post milking herbal teat dip disinfectant comprising of Berberis lycium, Curcuma longa and Eucalyptus globulus. The teat dip being aqueous in nature is not able to form long time barrier on teats leading to reappearance of mastitis-causing microbes. Discloses a herbal composition consisting of a synergistic combination of four essential oils viz. Cedrus deodara oil, Eucalyptus globulus oil, Curcuma longa oil and Gaultheria fragrantissima oil and one polysaccharide, preferably tamarind seed polysaccharide. The said composition upon drying makes a film around the teat which forms a long time barrier
7

at the teat canal opening and prevents microbes from penetrating into the teat canal.
No relevant invention was found in the Indian Patent Database.
OBJECTS OF THE PRESENT INVENTION
The object of the present invention is to disclose a topical barrier film-forming herbal composition
5 for the prevention and management of mastitis.
A further object is to disclose the method of preparation of the same.
Yet another object is to disclose the method of use of the composition.
A further object is to disclose the mechanism of action of the composition.
SUMMARY OF THE INVENTION
10 The present invention discloses a topical barrier film-forming herbal composition for the
prevention and management of mastitis. The said herbal composition consists of a synergistic combination of 4 essential oils viz. Cedrus deodara oil (1-3%, optimally 2%), Eucalyptus globulus oil (0.5-1.5%, optimally 1%), Curcuma longa oil (0.1-0.3%, optimally 0.2%), Gaultheria fragrantissima oil (0.5- 1.5%, optimally 1%) and a natural polysaccharide, preferably tamarind
15 seed polysaccharide (10-14%, optimally 12% (w/v). The composition shows high efficacy
(99.95%) against common mastitis-causing microbes. Furthermore, the composition not only has a preventive/ killing effect on microbes but also an emollient and wound-healing effect on skin wounds and fissures, thereby strengthening and enhancing the protection.
20 DETAILED DESCRIPTION OF THE DIAGRAMS
NIL
DETAILED DESCRIPTION OF THE INVENTION
The present invention discloses a topical barrier-film forming herbal composition for the
prevention and management of mastitis consisting of a synergistic combination of 4 essential oils
25 viz. Cedrus deodara oil (1-3%, optimally 2%), Eucalyptus globulus oil (0.5-1.5%, optimally 1%),
8

Curcuma longa oil (0.1-0.3%, optimally 0.2%), Gaultheria fragrantissima oil (0.5- 1.5%, optimally 1%) and a natural polysaccharide, preferably tamarind seed polysaccharide (10-14%, optimally 12% (w/v). The composition shows very high efficacy (99.5%) against common mastitis-causing microbes.
5 Method of preparation
The method of preparation of the composition is elaborated below and consists of the following steps:
Step 1- Preparation of aqueous phase
To hot water (80-90°C), a natural polysaccharide e.g. tamarind seed polysaccharide is
10 added in the concentration range of 10-14%, optimally 12% (w/v) and mixed properly to
prepare aqueous phase. The temperature of aqueous phase is lowered to 40-45°C and it is then stored.
Step 2 - Preparation of oil phase
The four oils viz. Cedrus deodara oil (1-3%, optimally 2%), Eucalyptus globulus oil (0.5-
15 1.5%, optimally 1%), Curcuma longa oil (0.1-0.3%, optimally 0.2%) and Gaultheria
fragrantissima oil (0.5- 1.5%, optimally 1%) are poured into a container and mixed. The
mixture of oils is then heated to 40-45°C. To this warm oil mixture, a suitable emollient
such as glycerin (2-3%, optimally 2.5%) and an anti-oxidant such as butylated hydroxyl
toluene (0.01%) are added with continuous stirring.
20 Step 3- Mixing of aqueous and oil phases
Aqueous and oil phases are mixed together in ratios 14:1 with vigorous shaking at 40-45°C.
The mixture is stirred for 10-20 minutes to get milky brown emulsion which represents
herbal composition of the present invention.
Evaluation of the herbal composition
25 Ten lactating, dairy cows were divided in two groups’ viz. control and test. The evaluation was
performed as given below:
Control group
The teats of the five cows, comprising the control group, were washed twice with sterile buffered normal saline at an interval of 2 hours and the buffer solution from each washing was then plated
9

separately onto Luria Bertani agar and incubated for 24 hours at 37 degrees C. The number of colony forming units (CFU) were counted.
Test group
The teats of the five cows, comprising the test group, were washed with sterile buffered normal
5 saline. The teats were then dipped in herbal composition of the present invention. The teat dip was
allowed to dry to form a topically adherent film and was allowed to remain on the teats for 2 hours;
thereafter, the film was peeled off and the teats were washed with sterile buffered normal saline.
This sterile buffered saline was cultured on Luria Bertani agar and incubated for 24 hours at 37
degrees C. The number of colony forming units (CFU) were counted.
10 Results
The results are summarized in Table 1
Table 1 Evaluation of herbal composition of the invention

S. no. Treatment of the teats of lactating animals CFU (colony forming units/ mL of normal saline) at 0 hrs CFU (colony forming units/ mL of normal saline) after 2 hrs Percent reduction in mastitis causing microbes
1. No treatment ~2 million ~2 million 0
2. Herbal composition ~2 million ~100 units only 99.95
These results compare well to currently available teat dips using harsh germicidal ingredients such
as iodine, but are safe and gentle and do not present any problem of irritation to either animals or
15 persons who come in contact with it or of contamination of the animals' milk supply.
Field Evaluation
The composition was also evaluated in the field, at dairy farms as below.
Methodology
30 healthy cows at onset of lactation were selected and divided into two equal groups:
20 Group I: Control animals (n = 15)
Teats were washed with water only as teat dip, twice daily immediately after each milking for a period of 45 days.
Group II: Test animals (n =15)
10

Teats were dipped with composition of present invention as teat dip, twice daily immediately after each milking for a period of 45 days.
Milk samples were collected from all animals before initiation of treatment at day 0 and post-
treatment on days 15, 30 and 45 and analyzed for Mean Somatic Cell Count (SCC). Higher SCC
5 in milk indicated ‘inflammation’ whereas low SCC indicated efficacy of the composition in
reducing infection/managing mastitis.
Results
Results are given in Table 2 and Table 3 below:
10 Table 2: Evaluation of efficacy of the composition in reducing Somatic Cell Count in milk

GROUPS SCC (×103)

0 day 15 days 30 days 45 days
Group I 108.92 ± 68.78 196.37 ± 110.28 222.38 ± 131.92 348.26 ± 188.48
Group II 111.18 ± 53.14 128.66 ± 59.74 114.24 ± 49.72 128.26 ± 55.26
Table 3: Evaluation of efficacy of the composition in reducing incidence of Mastitis

GROUPS Quarter-wise incidence (%) Animal-wise incidence (%)
Group I 17 (28.34%) 07 (46.67%)
Group II 03 (05.00%) 02 (13.34%)
15
Table 2 clearly shows that the composition of the present invention significantly prevents
mastitis in lactating animals. In the control group, Somatic Cell Count increases almost 3 fold
over a period of 45 days – from initial average count of about 108×103 to 348 ×103. In contrast,
animals treated with the composition of the present invention, show hardly any increase in the
20 Somatic Cell Count - count at beginning is 111×103 and at end of 45 days is 128 ×103.
Table 3 clearly shows that the composition of the present invention significantly reduces the incidence of mastitis. In the control group, the incidence of mastitis was 46% whereas in the treated group it was significantly less- only 13%.
Novelty, Inventive Step and Industrial Application
25 Novelty- The barrier film-forming herbal composition of present invention consisting of a
synergistic combination of 4 essential oils viz. Cedrus deodara oil, Eucalyptus globulus oil,
11

Curcuma longa oil, Gaultheria fragrantissima oil and a natural polysaccharide such as tamarind seed polysaccharide for the prevention and management of mastitis, has not been disclosed in any prior art and is novel.
Inventive step- The technical advancement of knowledge lies in disclosing a herbal composition for the treatment of mastitis which is equally effective as chemical based compositions but offers the technical advantages of being free from undesirable effects e.g. irritation and non-contamination of milk. Further, the composition is economical to manufacture as the essential oils required viz. Cedrus deodara oil, Eucalyptus globulus oil, Curcuma longa oil and Gaultheria fragrantissima oil and the polysaccharide added into the oils for film formation viz. tamarind seed polysaccharide, are easily and abundantly available.
Industrial application- The herbal composition can be easily manufactured at industrial scale by industries manufacturing veterinary medicines.
While the invention is described in detail in its preferred embodiments, it will be obvious to those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention e.g. minor changes in concentrations or reduction /addition of herbal extracts, use of different oils, polysaccharides from other sources apart from tamarind seed etc. Such changes and modifications shall be regarded as being within the scope of the invention.

claim:

1.A topical barrier film-forming herbal composition for prevention and management of mastitis WHEREIN the same consists of a synergistic combination of
i) Cedrus deodara oil- 1-3%, optimally 2%;
ii) Eucalyptus globulus oil- 0.5-1.5%, optimally 1%;
iii) Curcuma longa oil - 0.1-0.3%, optimally 0.2%;
iv) Gaultheria fragrantissima oil - 0.5- 1.5%, optimally 1%;
v) A natural polysaccharide - 10-14%, optimally 12% (w/v).

2. A herbal composition as claimed in claim 1 WHEREIN the natural polysaccharide present in the composition is tamarind seed polysaccharide.

3. A method for the preparation of the composition as claimed in claim 1 WHEREIN the same consists of the following steps:
Step 1- Preparation of aqueous phase
To hot water (80-90°C), a natural polysaccharide e.g. tamarind seed polysaccharide is added in the concentration range of 10-14%, optimally 12% (w/v) and mixed properly to prepare aqueous phase. The temperature of aqueous phase is lowered to 40-45°C and it is then stored.

Step 2 - Preparation of oil phase
The four oils viz. Cedrus deodara oil (1-3%, optimally 2%), Eucalyptus globulus oil (0.5-1.5%, optimally 1%), Curcuma longa oil (0.1-0.3%, optimally 0.2%) and Gaultheria fragrantissima oil (0.5- 1.5%, optimally 1%) are poured into a container and mixed. The mixture of oils is then heated to 40-45°C. To this warm oil mixture, a suitable emollient such as glycerin (2-3%, optimally 2.5%) and an antioxidant such as butylated hydroxyl toluene (0.01%) are added with continuous stirring.

Step 3- Mixing of aqueous and oil phases: Aqueous and oil phases are mixed together in ratios 14:1 with vigorous shaking at 40-45°C. The mixture is stirred for 10-20 minutes to get milky brown emulsion which represents herbal composition of the present invention.

KOMPAL BANSAL
PA NO. 1754
PATENT AGENT FOR THE APPLICANT
Dated: 31st March, 2018

, Description:FORM 2
THE PATENTS ACT, 1970
(39 OF 1970)
&
THE PATENTS RULES, 2003
COMPLETE SPECIFICATION
(SEE SECTION 10 AND RULE 13)

TOPICAL BARRIER FILM-FORMING HERBAL COMPOSITION FOR PREVENTION AND MANAGEMENT OF MASTITIS AND METHOD THEREOF

AYURVET LIMITED
VILLAGE KATHA, POST OFFICE BADDI,
DISTRICT SOLAN- 173205
HIMACHAL PRADESH, INDIA.
Email Id: sudheer@ayurvet.in
Contact no: 01795-247746

The following specification particularly describes the invention and the manner in which it is to be performed
FIELD OF INVENTION
The invention generally relates to the field of veterinary herbal products. More specifically, it relates to a topical barrier film-forming herbal composition for the prevention and management of mastitis and method thereof.
BACKGROUND OF THE INVENTION
Mastitis is the inflammation of the mammary gland and udder tissue and is a major endemic disease resulting in huge economic loss to the dairy industry. It usually occurs as an immune response to bacterial invasion of the teat canal by variety of bacterial species such as Staphylococcus aureus, Streptococcus faecalis, Streptococcus pyogenes, Klebsiella spp., Mycobacterium bovis and E. coli (Reference:https://dairy.ahdb.org.uk/technical-information/animal-healthwelfare/mastitis). The bacterial toxins released in the udder tissue damage the milk-secreting tissues and ducts throughout the udder. In some situations, the damage is permanent and the udder loses its functionality. Acute cases can lead to fatalities while cows that recover may be lactationally incapacitated for the rest of their lives.
Mastitis in cows can be a very complex problem since the farmer may not be able to detect it during its subclinical phase (a phase of mastitis without showing any signs and symptoms). At this stage, the disease can spread very fast to the rest of the herd if proper handling is lacking in the farm. The losses a farmer/ farm-owner is likely to incur arise from the following avenues:
• Milk thrown away due to contamination by medication or being unfit to drink.
• A reduction in yield due to illness and any permanent damage to udder tissue.
• The extra labor required to tend mastitic cows.
• The costs of veterinary care and medicines.
• The cost of reduced longevity due to premature culling.
Economic loss- Globally, mastitis accounts for about 38 per cent of the total direct cost of animal farming. In India, the economic losses due to mastitis have increased about 115-130 folds in last few years. Lack of awareness, delay in detection of mastitis, lack of markers for detecting ensuing mastitis, unhygienic milking practices, diverse production systems, inadequate treatment etc. are some of the important factors contributing to higher incidence of mastitis.
Existing methods for the prevention of mastitis and their limitations
The most common method for prevention of mastitis involves the use of antiseptic or antimicrobial agents, which are chemical in nature. These include iodophors, quaternary ammonium compounds, chlorhexidine salts, chlorine-release compounds (e.g. alkali hypochlorites), oxidizing compounds (e.g. hydrogen peroxide, peracids), protonated carboxylic acids (e.g. heptanoic, octanoic, nonanoic, decanoic, undecanoic acids), acid anionics (e.g. alkyl-aryl sulfonic acids), and chlorine dioxide (from chlorite). These agents have varying degrees of effectiveness and limit the transmission of mastitis by reducing pathogen populations on the teat surface.

Method of use- by dipping teats in solution of the compounds or spraying the solution on the teats. The solutions form a layer on the outer surface of the teats. The teat skin and muscles being tough, bacteria cannot enter through skin but only through the teat canal.
Limitations
i. Not efficient-due to insufficient amount of active agents or the insufficient contact time with the teat skin leading to incomplete prevention of mastitis.
ii. Recurrence of pathogens: It is also known that the effect of existing agents fades out very quickly and that renewed contamination of the teats takes place shortly after the treatment. Small wounds and skin tissues on the teats can act as reservoirs for certain microbes, and livestock with infected teat wounds often show higher mastitis frequencies than other livestock.
iii. Chemicals can enter into milk: Dipping of the teats into chemical solutions e.g. iodine, chlorhexidine and other agents can result in movement of these chemicals into milk which can not only be harmful for health but also interfere with the manufacture of dairy products.
In view of the foregoing, there exists a need for compositions which are safer and not harmful for health of animal or persons consuming milk from treated animals.

Introduction to present invention
The present invention discloses a topical barrier film-forming herbal composition for the prevention and management of mastitis. The said herbal composition consists of a synergistic combination of the following viz.
i. Cedrus deodara oil- 1-3%, optimally 2%
ii. Eucalyptus globulus oil- 0.5-1.5%, optimally 1%
iii. Curcuma longa oil - 0.1-0.3%, optimally 0.2%
iv. Gaultheria fragrantissima oil - 0.5- 1.5%, optimally 1%
v. A natural polysaccharide, preferably tamarind seed polysaccharide - 10-14%, optimally 12% (w/v).
Mechanism of action
When the said liquid composition is applied topically, it soon dries, forming a plug at the teat orifice, which acts like a mechanical barrier, thus preventing the entry of bacteria in the udder through the teat canal. Film formation also allows better surface retention of the active components on the surface of the teats. Further the synergistic mixture of oils results in broad-spectrum antimicrobial activity that kills nearly 99.95% of the common mastitis-causing bacteria. Further, the essential oils exert emollient (skin softening) and vulnerary (wound healing) activities on the teat skin.
Advantages
i. Safety- uses natural herbal ingredients and is thus free from the undesirable effects on human health associated with chemical compounds, which when applied on the teats, contaminate milk. The composition consists of ingredients that are classified under GRAS (Generally Regarded As Safe) category and, thus, eliminate the risks of contamination of milk.
ii. Unique mechanism of action- The herbal composition does not remain in ‘wet’ form on the teat after application, unlike the chemical compound solutions. It dries quickly, forming a protective barrier film, which prevents the entry of pathogens into the teat canal. Film formation allows better surface retention of the active components on the surface of the teats. The composition kills nearly all the common microbes, which may cause mastitis, due to broad spectrum bactericidal activity of the oils.
iii. Additional benefits - the composition not only has a preventive/killing effect on microbes but also a healing effect on skin wounds and fissures, thereby strengthening and enhancing the protection.
iv. Easy application- Furthermore, the composition can be given such consistency that it is easy to apply to the teats, for example by dipping or spraying, at the same time as it imparts an emollient effect on the teats. Also, the composition is not harsh on the milkers hand or milking machine and can be peeled off the teats or washed off easily with water before milking.
v. Economical production- All the ingredients viz. 4 essential oils and tamarind seed polysaccharide are easily available commercially and cheaply which makes production of the composition economical.
Prior art and limitations
S. No. Patent Application No. Prior Art Patent Present Invention
1. EP 0917427 Discloses a teat dip composition comprising of chitosan and a group of polysaccharides. The said composition could treat only 91% of the infected animals Consists of a synergistic combination of essential oils viz. Cedrus deodara oil, Eucalyptus globulus oil, Curcuma longa oil and Gaultheria fragrantissima oil and one polysaccharide, preferably tamarind seed polysaccharide in an optimum concentration. The composition of the present invention can prevent/ kill 99.95% of the common mastitis-causing microbes.
2. EP0904693 B1 Discloses a composition for treatment of mastitis which consists of an acidulant and chlorite. The said composition being chemical in nature can be harsh to the teats and hands of milker and can contaminate the milk.
Composition is gentle on the teat and free from any strong corrosive compounds. Rather it contains natural oils, which apart from antimicrobial effect, offer emollient and wound healing activities.
3. US 4945110 Discloses an antibacterial chemical composition for mastitis prevention which is an aqueous solution of an aliphatic sulfate or sulfonate salt detergent, lactic acid or a food grade salt. No chemical compound is present at all in composition of present invention.

4. US 5776479 Discloses a germicidal aqueous teat-dip composition which comprises a film-forming agent selected from the group consisting of hydroxyethylcellulose, methyl hydroxyl-propylcellulose, and ethylhydroxyethylcellulose, iodine complexed with a non-ionic surfactant and water. Composition of present invention is free from any synthetic film forming agents or chemical agents including non-ionic surfactants.
5. US 7078062 B2 Discloses an aqueous solution of hop-based udder and teat dips to wash and sanitize the teats. Composition is entirely different consisting of a synergistic mixture of 4 essential oils and a natural polysaccharide for film formation on drying.
6. US 2011/0086117 Al Discloses an aqueous antiseptic composition comprising only one essential oil i.e. 10 to 50% by volume of lemongrass oil and one solubilizing agent for solubilizing the lemongrass oil, 10 to 30% by volume of a carrier oil carrying the components of the composition and about 0.5 to 2% by volume of an electrolyte for providing conductivity for the com-position. The composition further comprises an organic acid for adjusting the pH level of the composition and an antioxidant. The composition is not aqueous but oil-in-water emulsion.
7. Waghmare et al., 2013. Int. J. Agric.Sc & Vet.Med. 41-50. Discloses an aqueous post milking herbal teat dip disinfectant comprising of Berberis lycium, Curcuma longa and Eucalyptus globulus. The teat dip being aqueous in nature is not able to form long time barrier on teats leading to reappearance of mastitis-causing microbes. Discloses a herbal composition consisting of a synergistic combination of four essential oils viz. Cedrus deodara oil, Eucalyptus globulus oil, Curcuma longa oil and Gaultheria fragrantissima oil and one polysaccharide, preferably tamarind seed polysaccharide. The said composition upon drying makes a film around the teat which forms a long time barrier at the teat canal opening and prevents microbes from penetrating into the teat canal.

No relevant invention was found in the Indian Patent Database.
OBJECTS OF THE PRESENT INVENTION
The object of the present invention is to disclose a topical barrier film-forming herbal composition for the prevention and management of mastitis.
A further object is to disclose the method of preparation of the same.
Yet another object is to disclose the method of use of the composition.
A further object is to disclose the mechanism of action of the composition.
SUMMARY OF THE INVENTION
The present invention discloses a topical barrier film-forming herbal composition for the prevention and management of mastitis. The said herbal composition consists of a synergistic combination of 4 essential oils viz. Cedrus deodara oil (1-3%, optimally 2%), Eucalyptus globulus oil (0.5-1.5%, optimally 1%), Curcuma longa oil (0.1-0.3%, optimally 0.2%), Gaultheria fragrantissima oil (0.5- 1.5%, optimally 1%) and a natural polysaccharide, preferably tamarind seed polysaccharide (10-14%, optimally 12% (w/v). The composition shows high efficacy (99.95%) against common mastitis-causing microbes. Furthermore, the composition not only has a preventive/ killing effect on microbes but also an emollient and wound-healing effect on skin wounds and fissures, thereby strengthening and enhancing the protection.

DETAILED DESCRIPTION OF THE DIAGRAMS
NIL
DETAILED DESCRIPTION OF THE INVENTION
The present invention discloses a topical barrier-film forming herbal composition for the prevention and management of mastitis consisting of a synergistic combination of 4 essential oils viz. Cedrus deodara oil (1-3%, optimally 2%), Eucalyptus globulus oil (0.5-1.5%, optimally 1%), Curcuma longa oil (0.1-0.3%, optimally 0.2%), Gaultheria fragrantissima oil (0.5- 1.5%, optimally 1%) and a natural polysaccharide, preferably tamarind seed polysaccharide (10-14%, optimally 12% (w/v). The composition shows very high efficacy (99.5%) against common mastitis-causing microbes.
Method of preparation
The method of preparation of the composition is elaborated below and consists of the following steps:
Step 1- Preparation of aqueous phase
To hot water (80-90°C), a natural polysaccharide e.g. tamarind seed polysaccharide is added in the concentration range of 10-14%, optimally 12% (w/v) and mixed properly to prepare aqueous phase. The temperature of aqueous phase is lowered to 40-45°C and it is then stored.
Step 2 - Preparation of oil phase
The four oils viz. Cedrus deodara oil (1-3%, optimally 2%), Eucalyptus globulus oil (0.5-1.5%, optimally 1%), Curcuma longa oil (0.1-0.3%, optimally 0.2%) and Gaultheria fragrantissima oil (0.5- 1.5%, optimally 1%) are poured into a container and mixed. The mixture of oils is then heated to 40-45°C. To this warm oil mixture, a suitable emollient such as glycerin (2-3%, optimally 2.5%) and an anti-oxidant such as butylated hydroxyl toluene (0.01%) are added with continuous stirring.
Step 3- Mixing of aqueous and oil phases
Aqueous and oil phases are mixed together in ratios 14:1 with vigorous shaking at 40-45°C. The mixture is stirred for 10-20 minutes to get milky brown emulsion which represents herbal composition of the present invention.
Evaluation of the herbal composition
Ten lactating, dairy cows were divided in two groups’ viz. control and test. The evaluation was performed as given below:
Control group
The teats of the five cows, comprising the control group, were washed twice with sterile buffered normal saline at an interval of 2 hours and the buffer solution from each washing was then plated separately onto Luria Bertani agar and incubated for 24 hours at 37 degrees C. The number of colony forming units (CFU) were counted.
Test group
The teats of the five cows, comprising the test group, were washed with sterile buffered normal saline. The teats were then dipped in herbal composition of the present invention. The teat dip was allowed to dry to form a topically adherent film and was allowed to remain on the teats for 2 hours; thereafter, the film was peeled off and the teats were washed with sterile buffered normal saline. This sterile buffered saline was cultured on Luria Bertani agar and incubated for 24 hours at 37 degrees C. The number of colony forming units (CFU) were counted.
Results
The results are summarized in Table 1
Table 1 Evaluation of herbal composition of the invention
S. no. Treatment of the teats of lactating animals CFU (colony forming units/ mL of normal saline) at 0 hrs CFU (colony forming units/ mL of normal saline) after 2 hrs Percent reduction in mastitis causing microbes
1. No treatment ~2 million ~2 million 0
2. Herbal composition ~2 million ~100 units only 99.95
These results compare well to currently available teat dips using harsh germicidal ingredients such as iodine, but are safe and gentle and do not present any problem of irritation to either animals or persons who come in contact with it or of contamination of the animals' milk supply.
Field Evaluation
The composition was also evaluated in the field, at dairy farms as below.
Methodology
30 healthy cows at onset of lactation were selected and divided into two equal groups:
Group I: Control animals (n = 15)
Teats were washed with water only as teat dip, twice daily immediately after each milking for a period of 45 days.
Group II: Test animals (n =15)
Teats were washed with composition of present invention as teat dip, twice daily immediately after each milking for a period of 45 days.
Milk samples were collected from all animals before initiation of treatment at day 0 and post-treatment on days 15, 30 and 45 and analyzed for Mean Somatic Cell Count (SCC). Higher SCC in milk indicated ‘infection’ whereas low SCC indicated efficacy of the composition in reducing infection/managing mastitis.
Results
Results are given in Table 2 and Table 3 below:

Table 2: Evaluation of efficacy of the composition in reducing Somatic Cell Count in milk
GROUPS SCC (×103)
0 day 15 days 30 days 45 days
Group I 108.92 ± 68.78 196.37 ± 110.28 222.38 ± 131.92 348.26 ± 188.48
Group II 111.18 ± 53.14 128.66 ± 59.74 114.24 ± 49.72 128.26 ± 55.26

Table 3: Evaluation of efficacy of the composition in reducing incidence of Mastitis
GROUPS Quarter-wise incidence (%) Animal-wise incidence (%)
Group I 17 (28.34%) 07 (46.67%)
Group II 03 (05.00%) 02 (13.34%)

Table 2 clearly shows that the composition of the present invention significantly prevents bacterial infection in milk from lactating animals. In the control group, the bacterial infection (as evidenced by enhanced Somatic Cell Count) increases almost 3 fold over a period of 45 days – from initial average count of about 108 to 348 (×103). In contrast, animals treated with the composition of the present invention, show hardly any increase in the cell count- count at beginning is 111 and at end of 45 days is 128 (×103).
Table 3 clearly shows that the composition of the present invention significantly reduces the incidence of mastitis. In the control group, the incidence of mastitis was 46% whereas in the treated group it was significantly less- only 13%.

Novelty, Inventive Step and Industrial Application
Novelty- The barrier film-forming herbal composition of present invention consisting of a synergistic combination of 4 essential oils viz. Cedrus deodara oil, Eucalyptus globulus oil, Curcuma longa oil, Gaultheria fragrantissima oil and a natural polysaccharide such as tamarind seed polysaccharide for the prevention and management of mastitis, has not been disclosed in any prior art and is novel.
Inventive step- The technical advancement of knowledge lies in disclosing a herbal composition for the treatment of mastitis which is equally effective as chemical based compositions but offers the technical advantages of being free from undesirable effects e.g. irritation and non-contamination of milk. Further, the composition is economical to manufacture as the essential oils required viz. Cedrus deodara oil, Eucalyptus globulus oil, Curcuma longa oil and Gaultheria fragrantissima oil and the polysaccharide added into the oils for film formation viz. tamarind seed polysaccharide, are easily and abundantly available.
Industrial application- The herbal composition can be easily manufactured at industrial scale by industries manufacturing veterinary medicines.

While the invention is described in detail in its preferred embodiments, it will be obvious to those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention e.g. minor changes in concentrations or reduction /addition of herbal extracts, use of different oils, polysaccharides from other sources apart from tamarind seed etc. Such changes and modifications shall be regarded as being within the scope of the invention.

KOMPAL BANSAL
PA NO. 1754
PATENT AGENT FOR THE APPLICANT
Dated: 31st March, 2018