Nipple sealant composition and method of preventing mastitis

Abstract

A teat sealant composition free of bismuth salts, a method for preparing the same, and a method for preventing new intra-breast infections or mastitis.

Description

[0001] Related applications

[0002] This application claims priority to Australian patent application number 2023900582 filed on 6 March 2023, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to teat sealant compositions that do not contain bismuth salts, methods for their preparation, and methods for preventing new intramammary infections or mastitis.

[0004] background

[0005] In dairy cows, the incidence of mastitis caused by environmental pathogens is highest at the beginning of the 7-10 week dry period and again at calving at the end of the dry period. During the dry period, cows develop a natural plug, the keratin plug, which seals the teat canal; however, this takes some time (several weeks), during which microorganisms can enter and cause mastitis.

[0006] Internal teat sealants (ITS) containing heavy metal bismuth salts are used during the dry period of dairy cows and heifers to "seal" the teat canals and thereby prevent, or at least reduce, the entry of environmental pathogens into the udder. Teat sealants can be used alone or in combination with intramammary antibiotics (dry cow therapy). Dry cow therapy containing antibiotics has been used for many years to reduce the incidence of mastitis in dairy cows during lactation.

[0007] Many studies have been published on the effectiveness of ITS in preventing new intramammary infections during the dry period and early lactation mastitis. If combined with antibiotic dry cow therapy, they show additional effectiveness. Most countries adopt a targeted treatment approach, in which cows with dry period (late lactation) infection receive antibiotic therapy to cure the existing infection and then also receive non-antibiotic teat sealants to prevent new infections during the dry period; while cows that are not infected during the dry period simply receive non-antibiotic teat sealants.

[0008] Growing concerns about the widespread use of antimicrobials in food-producing animals have shifted attitudes toward dry-cow therapy. Consequently, the use of non-antibiotic teat sealants is increasing. Most ITS use bismuth subnitrate as their primary component, i.e., up to 65% w / w. Consumers may perceive bismuth subnitrate as an undesirable heavy metal in milk. Milk processors in New Zealand have recently become so concerned about high levels of bismuth residues that they are considering reducing the dosage of commercially available products.

[0009] The increased uptake of bismuth-based products has led to an increase in bismuth residues in milk and milk products, which is currently under scrutiny by regulators and milk processors. However, due to limited alternatives and ongoing pressure to reduce antibiotic use, there are no alternatives to bismuth-based sealants. Detailed Description of the Invention

[0011] It is an object of one or more embodiments of the present invention to provide a teat sealant composition that does not contain bismuth salts (ie, is free of bismuth salts) for preventing new intramammary infections or mastitis, or to otherwise provide the public with a useful commercial choice.

[0012] Composition

[0013] According to a first aspect of the present invention, there is provided a teat sealant composition free of bismuth salts, formulated for occluding teat canals and / or teat cistern.

[0014] According to a second aspect of the present invention there is provided a teat sealant composition comprising a salt other than a bismuth salt, formulated for occluding a teat duct and / or teat cistern.

[0015] According to a third aspect of the present invention, there is provided a teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle, wherein the solid particles are not a bismuth salt.

[0016] According to a fourth aspect of the present invention, there is provided a teat sealant composition comprising solid particles in the form of a gel, a paste or a coarse suspension, which is formulated for occluding teat ducts and / or teat cistern.

[0017] Preferably, the composition is free of bismuth salts and bismuth compounds.

[0018] Preferably, the composition is formulated for administration to and retention in the teat duct and / or teat cistern of a cow.

[0019] Preferably, the composition is in the form of a gel, paste or coarse suspension.

[0020] Preferably, the composition has a sufficiently low viscosity to allow it to be administered by injection through the teat canal and / or teat cistern, preferably using a syringe. Preferably, the composition can be administered to the teat using a syringe (a syringe utilizing a plunger), preferably an intramammary syringe.

[0021] Any suitable amount may be administered. Typically, about 2-5 g of the composition is administered / injected.

[0022] Preferably, the injection force for injecting the composition into the nipple does not exceed 4500 g at 25° C. Preferably, the injection force for injecting the composition into the nipple at 25° C. is about 1500 g to 3500 g, including all values ​​between the upper and lower limits and any subranges falling within the range (including about 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3200, 3200, 3300, 3400, 3500). Please note that this force depends not only on the formulation of the composition, but also on the design of the syringe. Preferably, the syringe is a standard intramammary syringe with a collar / nozzle inner diameter of 1.5 mm, an outer diameter of 2.5 mm, an inner barrel diameter of 13.5 mm, a collar / nozzle length of 12 mm, and a barrel length of 49 mm. Such syringes are sold by Hubert De Backer nv (HDB Veterinary Intramammary Syringes) at https: / / www.hdb.be / _library / _files / Tabellen / 0.5.0 / Intramammary_0_5_0.pdf. Unless otherwise noted, the injectability studies described herein utilized this HDB Veterinary Intramammary Syringe. Preferably, the composition thickens rapidly in the teat, thereby being retained and occluding the teat duct and / or teat cistern.

[0023] Typically, the clumps of composition in the teat canal are very small compared to the clumps of composition in the teat cistern.

[0024] Preferably, once located within the teat canal and / or teat cistern (or at least partially within the teat cistern), the composition has a sufficiently high viscosity to remain within the teat canal or teat cistern and to occlude the teat canal or teat cistern.

[0025] Preferably, the composition has a rebound viscosity such that it can be retained within the teat canal and / or teat cistern as a substantially cohesive mass, wherein "rebound" means that the viscosity increases fairly quickly once the sealant is delivered into the teat.

[0026] Preferably, the composition has specific rheological properties that enable the composition to be administered into the teat duct and / or teat cistern by injection and to be retained in the teat duct and / or teat cistern and to block the teat duct and / or teat cistern or to prevent intramammary infection.

[0027] Preferably, the composition has a viscosity greater than about 1 g / cm 3 The density is preferably about 1.3 g / cm 3 Up to 1.6g / cm 3between (approximately 1.3, 1.4, 1.5 and 1.6 inclusive).

[0028] The phrase "formulated to occlude" means that the composition is capable of forming a physical barrier in the teat duct and / or teat cisterns, thereby preventing or substantially preventing pathogenic microorganisms, such as bacteria, from entering the udder.

[0029] Preferably, the composition is shear thinning under the shear forces applied during injection and expression from the nipple.

[0030] In certain embodiments, the composition has antimicrobial activity, particularly antibacterial activity, particularly activity against bacteria that cause mastitis. The antibacterial activity may be antibacterial or bactericidal.

[0031] In certain embodiments, the composition occludes the teat ducts and / or teat cistern for at least about 14 days, typically about 42 to about 90 days (including 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, and 90 days and all subranges therebetween), or typically for a period of up to about 4 months (including about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, and 16 weeks).

[0032] In certain embodiments, the obstruction persists throughout the dry period.

[0033] In certain embodiments, blocking continues for approximately the first 14 days of the dry period, until a natural keratin plug forms.

[0034] The composition is preferably used in dairy cows or heifers.

[0035] In certain embodiments, the composition occludes or substantially occludes the teat duct and / or teat cistern until a natural keratin plug has formed in the teat duct.

[0036] In certain embodiments, the composition occludes or substantially occludes the teat ducts and / or teat cisterns for about 4-6 weeks while allowing a natural keratin plug to form.

[0037] In certain embodiments, the composition is used when the cow is drying off.

[0038] In certain embodiments, the composition is used when environmental pathogens are greatest, ie, at the beginning of the dry period.

[0039] In certain embodiments, the composition is formulated such that no milking from the teat canal and / or teat cistern is required at the end of the dry period.

[0040] All ingredients of the composition may be veterinarily acceptable.

[0041] Preferably, some, most or all of the ingredients of the composition are biodegradable or environmentally safe.

[0042] In certain embodiments, the composition is antimicrobial or antibacterial, including inhibiting the growth of microorganisms or killing microorganisms (e.g., killing bacteria). In certain embodiments, one or more components of the composition can provide antimicrobial activity. In certain embodiments, the solid particles can change the rheological properties of the composition and provide antimicrobial activity.

[0043] The composition may have antimicrobial activity against bacteria that cause mastitis. The composition may have antimicrobial activity against any one or more of the following bacteria: Streptococcus uberis, Staphylococcus aureus, and Escherichia coli (E. coli).

[0044] The solid particles may be very soluble, freely soluble, soluble, sparingly soluble, slightly soluble, very slightly soluble, practically insoluble or insoluble relative to water. Guidelines for water solubility levels are defined as follows (and are not to be considered as fixed, exact numbers):

[0045]

[0046] Advantages of using water-soluble solid particles include that they can reduce the amount of residue left in the teat canal (and udder) that needs to be squeezed out or filtered out during milking; and that some water-soluble solid materials can provide an antimicrobial effect around the teat sealant composition by forming a hypertonic solution.

[0047] Any suitable type or types of solid particles (excluding heavy metal bismuth salts) may be used in any suitable amount.

[0048] In certain embodiments, the solid particles have a solubility in water exceeding about two times the isotonic concentration.

[0049] In certain embodiments, the solid particles comprise at least one type of non-bismuth salt, such as an organic salt or an inorganic salt. These include, but are not limited to, halide salts, phosphates, and carbonates, such as sodium chloride (which is water-soluble), potassium chloride (which is water-soluble), calcium phosphate (which is very slightly soluble, almost insoluble, or insoluble in water, depending on its form), and magnesium carbonate (which is slightly soluble in water). The solid particles may comprise at least one type of heavy metal salt, provided that it is not a bismuth salt and does not produce toxic or undesirable metal residues in the milk or meat of the animal. Acceptable heavy metals may include calcium, magnesium, and aluminum, but do not include toxic heavy metals.

[0050] The term "calcium phosphate" as defined herein includes within its scope any calcium phosphate containing calcium phosphate, if the context permits. 3 - 4. HPO 2- 4 or H2PO - 4 combinations of Ca 2+ Compounds of the invention, including anhydrous and hydrate forms (salt hydrates), include: monocalcium phosphate; dicalcium phosphate; tricalcium phosphate (which is practically insoluble or insoluble in water); anhydrous dibasic calcium phosphate (which is very slightly soluble) / anhydrous dicalcium phosphate; dibasic calcium phosphate dihydrate (which is very slightly soluble) / dicalcium phosphate dihydrate / dibasic calcium phosphate dihydrate; anhydrous monocalcium phosphate / anhydrous bis(monobasic) calcium phosphate / anhydrous monocalcium phosphate; monobasic calcium phosphate monohydrate / bis(monobasic) calcium phosphate monohydrate / monobasic calcium phosphate monohydrate; and monobasic calcium phosphate dihydrate (which is practically insoluble or very slightly soluble in water) / dicalcium phosphate dihydrate / dibasic calcium phosphate dihydrate.

[0051] In certain embodiments, the solid particles comprise at least one type of carbohydrate, such as a simple sugar or a complex sugar. These include, but are not limited to, sucrose (which is very soluble in water) and mannitol (which is readily soluble in water).

[0052] In certain embodiments, the solid particles comprise at least one type of mineral, such as a clay, such as kaolin (which is generally insoluble in water), such as kaolinite, halloysite, dickite, or nacrite.

[0053] In certain embodiments, the solid particles comprise sodium chloride. In certain embodiments, the solid particles comprise potassium chloride. In certain embodiments, the solid particles comprise calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / dicalcium phosphate dihydrate). In certain embodiments, the solid particles comprise calcium carbonate. In certain embodiments, the solid particles comprise magnesium carbonate.

[0054] In certain embodiments, the solid particles comprise sodium chloride or potassium chloride and calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate). In certain embodiments, the solid particles comprise sodium chloride or potassium chloride and calcium carbonate. In certain embodiments, the solid particles comprise sodium chloride or potassium chloride and magnesium carbonate.

[0055] In certain embodiments, the composition comprises solid particles in the following approximate amount ranges: 10% to 75%, 35% to 70%, 40% to 65%, 50% to 60%, w / w, including all values ​​between 10 and 75, including about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75 8, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74 and 75.

[0056] In some embodiments, the solid particles comprise approximately 25-60% w / w sodium chloride or potassium chloride (including approximately 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, and 60).

[0057] In certain embodiments, the solid particles comprise about 10-30% w / w magnesium carbonate (including about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30).

[0058] In certain embodiments, the solid particles comprise about 10% w / w magnesium carbonate and about 25% w / w sodium chloride or potassium chloride.

[0059] In certain embodiments, the solid particles comprise approximately 55% w / w sodium chloride or potassium chloride.

[0060] In certain embodiments, the solid particles comprise approximately 35-60% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate) (including approximately 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 and 60).

[0061] In certain embodiments, the solid particles comprise approximately 60% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate).

[0062] In certain embodiments, the solid particles comprise about 10% w / w sodium chloride or potassium chloride and about 50% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate).

[0063] In certain embodiments, the solid particles comprise approximately 25% w / w sodium chloride or potassium chloride and approximately 35% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate).

[0064] In certain embodiments, the solid particles comprise about 10% w / w sodium chloride or potassium chloride and about 50% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate).

[0065] In some embodiments, the solid particles can be ground and / or sieved. In some embodiments, the solid particles can be within a range of particle sizes, or have a particular average particle size. In some embodiments, the average particle size can be less than about 110, 109, 108, 107, 106, 105, 104, 103, 102, 101, 100, 99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82, 81, 80, 79, 78, 77, 76, 75, 74, 73, 72, 71, 70, 69, 68, 67, 66, 65, 64, 63 , 62, 61, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, 50, 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, or 3 μm in size. In certain embodiments, the average particle size may be less than about 106 μm in size. In certain embodiments, the average particle size may be less than about 80 μm in size. In certain embodiments, the average particle size may be less than about 50 μm in size. In certain embodiments, the average particle size can be less than about 30 μm in size. In certain embodiments, the average particle size can be less than about 20 μm in size. In certain embodiments, the average particle size can be less than about 10, 15, 20, 25, 30, 35 or 40 μm in size. In certain embodiments, the average particle size can be about 10-40 μm in size (including about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 and 40). In certain embodiments, the average particle size can be about 10-50 μm in size (including about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50). In certain embodiments, the average particle size can be about 20-30 μm in size (including about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30).In certain embodiments, the average particle size may be about 1-10 μm in size (including about 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10). In certain embodiments, the average particle size may be greater than about 3 μm in size.

[0066] In certain embodiments, D90 (90% particle size distribution parameter) can be about 30-60 μm (including about 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, and 60); D50 can be about 6-12 μm in size (including about 6, 7, 8, 9, 10, 11, and 12); and / or D10 can be 1-2 μm. In certain embodiments, these D90, D50, and D10 particle size distribution parameters are applicable to sodium chloride. In certain embodiments, these D90, D50, and D10 particle size distribution parameters are applicable to calcium phosphate as defined herein, preferably dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate. In certain embodiments, the D90 of calcium phosphate as defined herein may be about 50 μm. In certain embodiments, the sodium chloride may be ground to an average particle size of about <10 μm. In certain embodiments, the sodium chloride may have an average particle size of about 20-30 μm.

[0067] Any suitable type of vehicle can be used. In certain embodiments, the vehicle can include a water-insoluble carrier and a thickener. Typically, the thickener comprises solid particles. That is, the solid particles typically affect the rheological properties of the composition.

[0068] Any suitable type of water-insoluble carrier can be used. The water-insoluble carrier can include at least one type of carrier agent. In certain embodiments, the carrier or carrier agent includes at least one type of oil or oily liquid. Suitable examples include vegetable oils, mineral oils, synthetic oils, medium chain triglycerides and triglycerides. Preferred examples include (light or heavy) paraffin oil, liquid paraffin, (white or yellow) petrolatum, sesame oil and miglyol (which is a medium chain triglyceride).

[0069] Any suitable amount of carrier or carrier agent can be used. In certain embodiments, the composition comprises a carrier or carrier agent in the approximate range of 20% to 80% w / w (including about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, and 65).

[0070] Any suitable type of thickener can be used. The thickener can include at least one type of thickening agent. The at least one type of thickening agent can be water-soluble and / or water-insoluble. In certain embodiments, at least one thickening agent can function as both a thickener and an antimicrobial agent. An example of such an agent is lauric acid.

[0071] In certain embodiments, the at least one thickening agent can comprise a medium chain saturated fatty acid, such as lauric acid. In certain embodiments, lauric acid is used.

[0072] In certain embodiments, the composition comprises approximately 1% to 10% w / w medium chain saturated fatty acids, such as lauric acid, including all numbers between 1 and 10 (including approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10), including approximately 1%, 2.5%, and 5%.

[0073] In certain embodiments, at least one thickening agent can comprise a metal salt of a long-chain fatty acid, such as stearate. Suitable salts can include aluminum distearate, aluminum stearate, aluminum tristearate, ammonium stearate, barium stearate, butyl stearate, cadmium stearate, calcium stearate, cobalt stearate, copper stearate, ethylene glycol stearate, lithium stearate, magnesium stearate, manganese stearate, methyl stearate, potassium stearate, sodium stearate, strontium stearate and zinc stearate. Preferably, at least one thickening agent comprises aluminum stearate.

[0074] In certain embodiments, the composition comprises about 1% to 10% w / w metal salts of long chain fatty acids (including about 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10), such as stearates, such as aluminum stearate, including all numbers between 1 and 10 (including about 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10), including about 4.3 and 5.3%.

[0075] In certain embodiments, at least one thickening agent may comprise silica, such as hydrophilic or hydrophobic silica. Suitable silica agents include fumed silica, such as hydrophilic fumed silica or hydrophobic fumed silica. Hydrophilic fumed silica is sold under the trademark Aerosil-200. Hydrophobic fumed silica is sold under the trademark Aerosil R972.

[0076] In certain embodiments, the composition comprises approximately 0.1% to 1% w / w silica, such as hydrophilic fumed silica, including all values ​​between 0.1 and 1, including approximately 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, and 1, including approximately 0.7%.

[0077] In certain embodiments, the at least one thickening agent may comprise water-soluble fumed silica, such as the product sold by Evonik under the trademark Aerosil-200.

[0078] Any suitable amount of thickener or thickening agent can be used. In certain embodiments, the composition comprises a thickener or thickening agent in the approximate range of 0.1% to 10%, w / w, including all values ​​between 0.1 and 10 (including approximately 0.1, 0.2 ... 1, 1.1, 1.2 ... 2, 2.1, 2.2 ... 3, 3.1, 3.2 ... 4, 4.1, 4.2 ... 5, 5.1, 5.2 ... 6, 6.1, 6.2 ... 7, 7.1, 7.2 ... 8, 8.1, 8.2 ... 9, 9.1, 9.2 ... 10), including approximately less than 1%.

[0079] The composition may have antimicrobial activity against bacteria that cause mastitis.The composition may have antimicrobial activity, for example, against any one or more of the following bacteria: Streptococcus uberis, Staphylococcus aureus and / or Escherichia coli (E. coli).

[0080] In certain embodiments, the composition or its components, agents or ingredients can provide contact killing. In certain embodiments, the composition or its components, agents or ingredients can provide solution killing. In certain embodiments, the composition or its components, agents or ingredients can provide contact killing and solution killing.

[0081] "Contact kill" means that the surface of the composition has antimicrobial activity when surface contact with bacteria, etc. "Solution kill" means that the chemical, ingredient or agent leaches from the composition and has antimicrobial activity when in contact with bacteria, etc.

[0082] In certain embodiments, the composition comprises at least one type of antimicrobial, particularly an antibacterial. Suitable agents include chlorhexidine, PVP-iodine, nisin, and lauric acid. Preferably, the composition does not contain antibiotics. "Antibiotic" in this context refers to a class of antibiotics commonly / conventionally used in dry cow therapy. Any suitable amount of antimicrobial can be used. In certain embodiments, the composition comprises an antimicrobial in the following approximate range: 0.1% to 10%, w / w, including all values ​​between 0.1 and 10 (including approximately 0.1, 0.2...1, 1.1, 1.2...2, 2.1, 2.2...3, 3.1, 3.2...4, 4.1, 4.2...5, 5.1, 5.2...6, 6.1, 6.2...7, 7.1, 7.2...8, 8.1, 8.2...9 , 9.1, 9.2…10), including about 1-5% (including about 1, 2, 3, 4, and 5) for antimicrobial agents such as lauric acid, and about 0.2-2% (including about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, and 2) for antimicrobial agents such as chlorhexidine.

[0083] In certain embodiments, the antimicrobial activity is provided by the vehicle. In certain embodiments, the antimicrobial activity is provided by the solid particles. In certain embodiments, the antimicrobial activity is provided by both the vehicle and the solid particles. In certain embodiments, the antimicrobial activity is provided by a component of the vehicle (such as a carrier or a carrier agent thereof and / or a thickener or a thickening agent thereof). For example, lauric acid can be a thickening agent and can provide antibacterial activity. For example, a salt such as sodium chloride can be a thickening agent and can provide antibacterial activity by forming a hypertonic solution around the composition in the teat duct and / or teat cistern.

[0084] The composition may include one or more other ingredients such as at least one antioxidant or preservative. Any suitable amount may be added, such as from about 0.01% w / w to 5% w / w (including about 0.01...0.05...0.1...0.5...2...2.5...3...3.5...4...4.5...5). Examples of antioxidants or preservatives include alpha tocopherol, ascorbic acid, ascorbyl palmitate, benzalkonium chloride, benzethonium chloride, benzoic acid, benzyl alcohol, BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene), bronopol, butylparaben, cetyltrimethylammonium bromide, chlorhexidine, chlorobutanol, chlorocresol, cresol, ethylparaben, fumaric acid, imidurea, malic acid, monothioglycerol, n-propyl gallate, parabens (methylparaben and / or propylparaben), phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric acetate, phenylmercuric borate, phenylmercuric nitrate, potassium sorbate, sodium ascorbate, sodium benzoate, sodium metabisulfite, sodium propionate, sorbic acid, and thimerosal.

[0085] The composition may include at least one colorant. Any suitable amount may be added, such as from about 0.01% w / w to 10% w / w (including about 0.01 ... 0.1 ... 1 ... 10). Examples of colorants include dyes and pigments, including aluminum lakes, caramel, methylene blue, and titanium dioxide.

[0086] According to a fifth aspect of the present invention, there is provided a nipple sealant composition comprising:

[0087] an oily carrier; and

[0088] a thickener comprising solid particles and lauric acid,

[0089] wherein the solid particles are not bismuth metal salts.

[0090] According to a sixth aspect of the present invention there is provided a nipple sealant composition comprising a coarse filler material in the form of solid particles in a gelling agent, wherein the solid particles are not a bismuth metal salt.

[0091] According to a seventh aspect of the present invention, there is provided a nipple sealant composition comprising:

[0092] Liquid paraffin;

[0093] Aluminum stearate;

[0094] fumed silica; and

[0095] Water-soluble solid particles,

[0096] The composition is in the form of a paste or a suspension.

[0097] According to an eighth aspect of the present invention, there is provided a nipple sealant composition comprising:

[0098] Liquid paraffin;

[0099] Aluminum stearate;

[0100] fumed silica; and

[0101] Water-insoluble solid particles,

[0102] The composition is in the form of a paste or a suspension.

[0103] In certain embodiments, the teat sealant composition comprises one of the following formulations shown in the formulation table (all ingredients shown are actual or approximate values ​​and are in % w / w).

[0104] Preparation table.

[0105]

[0106]

[0107] Preparation method

[0108] According to a ninth aspect of the present invention, there is provided a method for preparing a teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle, the method comprising the steps of:

[0109] (1) forming a water-insoluble gel by mixing at least one type of water-insoluble carrier agent with at least one type of thickening agent;

[0110] (2) adding solid particles to the gel to form a paste; and, optionally,

[0111] (3) adding at least one other type of thickening agent to the gel or paste, wherein:

[0112] preparing a teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle;

[0113] The solid particles are not bismuth metal salts;

[0114] Step (3) is optional; and

[0115] Steps (2) and (3) do not need to be performed in the order described.

[0116] According to a tenth aspect of the present invention, there is provided a method for preparing a nipple sealant composition, the method comprising the following steps:

[0117] (1) forming a water-insoluble gel by mixing at least one type of oily vehicle with at least one type of thickening agent;

[0118] (2) adding solid particles to the gel to form a paste; and,

[0119] (3) adding lauric acid as a thickening agent to the gel or paste, wherein:

[0120] The solid particles are not bismuth metal salts.

[0121] According to an eleventh aspect of the present invention, there is provided a teat sealant composition prepared by the method according to the ninth or tenth aspect of the present invention.

[0122] Unless otherwise stated, "cow" as used in this specification encompasses "heifer".

[0123] How to use

[0124] According to a twelfth aspect of the present invention there is provided a teat sealant composition for preventing new intramammary infection or mastitis in a cow, the composition being as described in relation to the first to eighth and eleventh aspects of the present invention.

[0125] According to a thirteenth aspect of the present invention there is provided a teat sealant composition for use in preventing new intramammary infection or mastitis in a cow, or when used for such use, the composition being as described in relation to the first to eighth and eleventh aspects of the present invention.

[0126] According to a fourteenth aspect of the present invention there is provided use of a teat sealant composition in the preparation of a medicament for preventing new intramammary infection or mastitis in a cow, the composition being as described in relation to the first to eighth and eleventh aspects of the present invention.

[0127] According to a fifteenth aspect of the present invention, there is provided a method for preventing new intramammary infection or mastitis in a cow, the method comprising the step of applying to at least one teat duct and / or teat cistern of the cow a composition as described in relation to the first to eighth and eleventh aspects of the present invention.

[0128] According to a sixteenth aspect of the present invention, there is provided use of the composition described in relation to the first to eighth and eleventh aspects of the present invention for preventing new intramammary infection or mastitis in a cow.

[0129] According to a seventeenth aspect of the present invention, there is provided a syringe containing the composition described in relation to the first to eighth and eleventh aspects of the present invention.

[0130] According to an eighteenth aspect of the present invention, there is provided a kit for preventing new intramammary infection or mastitis in a cow or when used in a method of preventing new intramammary infection or mastitis in a cow, wherein the kit comprises: a syringe capable of administering a composition as described in relation to the first to eighth and eleventh aspects of the present invention to at least one teat canal and / or teat cistern of a cow.

[0131] The composition is preferably used in dairy cows or heifers.

[0132] Preferably, the composition prevents new intramammary infection or mastitis for at least about 14 days, typically about 42 to about 90 days (including 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105 , 83, 84, 85, 86, 87, 88, 89 and 90 days and all subranges therebetween), or generally for a period of up to about 4 months (including about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5 and 16 weeks).

[0133] In certain embodiments, the composition achieves blocking for approximately the first 14 days of the dry period, until a natural keratin plug forms.

[0134] In certain embodiments, the composition occludes or substantially occludes the teat duct and / or teat cistern until a natural keratin plug has formed in the teat duct.

[0135] Preferably, the composition protects the cow's teat or teats from bacterial infection throughout the dry period.

[0136] Preferably, the composition is administered per quarter at the end of lactation.

[0137] Preferably, the composition is administered per quadrant in primiparous heifers approximately 4 weeks prior to calving.

[0138] Preferred embodiments of the invention are defined in the following numbered paragraphs.

[0139] 1. A teat sealant composition free of bismuth salts, formulated for occluding teat ducts and / or teat cistern.

[0140] 2. A teat sealant composition comprising a salt other than a bismuth salt, formulated for occluding the teat duct and / or teat cistern.

[0141] 3. A teat sealant composition formulated for application to and retention within the teat canal and / or teat cistern of a cow.

[0142] 4. The teat sealant composition of any of the preceding paragraphs, wherein the teat sealant composition is in the form of a gel, paste, or coarse suspension.

[0143] 5. The teat sealant composition of any of the preceding paragraphs, wherein:

[0144] - the teat sealant composition has a sufficiently low viscosity to allow it to be administered by injection through the teat canal and / or teat cistern;

[0145] - Apply / inject approximately 2-5g of the teat sealant composition into the teat canal and / or teat cistern;

[0146] - the injection force for injecting the teat sealant composition into the teat at 25°C does not exceed 4500g, and preferably the injection force for injecting the teat sealant composition into the teat at 25°C is between about 1500g and 3500g;

[0147] - the teat sealant composition rapidly thickens in the teat, thereby being retained and blocking the teat ducts and / or teat cistern;

[0148] - once located within the teat canal and / or teat cistern, the teat sealant composition has a sufficiently high viscosity to be retained within and block the teat canal or teat cistern;

[0149] - the teat sealant composition has a rebound viscosity enabling it to be retained within the teat canal and / or teat cistern as a substantially cohesive mass;

[0150] The teat sealant composition has rheological properties that enable the composition to be administered into the teat duct and / or teat cistern by injection and to be retained in the teat duct and / or teat cistern to block the teat duct and / or teat cistern or to prevent intramammary infection;

[0151] - The teat sealant composition has a viscosity greater than about 1 g / cm 3 The density is preferably about 1.3 g / cm 3 Up to 1.6g / cm 3 between;

[0152] - the teat sealant composition shear thins under the shear forces applied during injection and milking from the teat;

[0153] - the teat sealant composition has antimicrobial activity, preferably antibacterial activity, preferably against bacteria that cause mastitis;

[0154] - the teat sealant composition is formulated to occlude the teat duct and / or teat cistern for a period of at least about 14 days, preferably about 42 to about 90 days;

[0155] - the teat sealant composition is formulated to occlude the teat duct and / or teat cistern throughout the dry period;

[0156] - the teat sealant composition is formulated to occlude the teat ducts and / or teat cisterns for at least about the first 14 days of the dry period until a natural keratin plug forms;

[0157] - the teat sealant composition occludes or substantially occludes the teat ducts and / or teat cistern for about 4-6 weeks while allowing the natural keratin plug to form; or

[0158] - The teat sealant composition is formulated so that it does not need to be milked from the teat canal and / or teat cistern at the end of the dry period.

[0159] 6. A teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle, wherein the solid particles are not a bismuth salt.

[0160] 7. The teat sealant composition of paragraph 6, wherein: the antimicrobial activity is provided by the vehicle; the antimicrobial activity is provided by the solid particles; the antimicrobial activity is provided by both the vehicle and the solid particles; the antimicrobial activity is provided by a component of the vehicle; or the composition further comprises at least one type of antimicrobial agent, preferably an antibacterial agent.

[0161] 8. The teat sealant composition of paragraph 6 or paragraph 7, wherein the vehicle comprises a water-insoluble carrier and a thickener, and preferably the thickener comprises the solid particles.

[0162] 9. The teat sealant composition of paragraph 8, wherein the water-insoluble carrier comprises at least one type of carrier agent, and the carrier agent comprises at least one type of oil or oily liquid.

[0163] 10. The teat sealant composition of paragraph 9, wherein the at least one type of oil or oily liquid comprises vegetable oil, mineral oil, synthetic oil, medium chain triglyceride or triglyceride, preferably paraffin oil, liquid paraffin, petrolatum, sesame oil or medium chain triglyceride.

[0164] 11. The teat sealant composition of paragraph 9 or 10, wherein the composition comprises approximately 20% to 80% w / w or 30-65% w / w of at least one type of carrier agent.

[0165] 12. The teat sealant composition of any of paragraphs 8 to 11, wherein the thickener comprises at least one type of water-soluble and / or water-insoluble thickening agent, preferably in an amount of about 0.1% to 10% w / w.

[0166] 13. The teat sealant composition of paragraph 12, wherein the at least one thickening agent functions as both a thickening agent and an antimicrobial agent.

[0167] 14. The teat sealant composition of paragraph 12 or paragraph 13, wherein the at least one thickening agent comprises a medium chain saturated fatty acid, preferably lauric acid.

[0168] 15. The teat sealant composition of paragraph 14, wherein the composition comprises about 1% to 10% w / w medium chain saturated fatty acid, preferably lauric acid.

[0169] 16. The teat sealant composition of paragraph 12 or 13, wherein the at least one thickening agent comprises a metal salt of a long chain fatty acid, preferably a stearate.

[0170] 17. The nipple sealant composition of paragraph 16, wherein the metal salt of a long-chain fatty acid comprises aluminum distearate, aluminum stearate, aluminum tristearate, ammonium stearate, barium stearate, butyl stearate, cadmium stearate, calcium stearate, cobalt stearate, copper stearate, ethylene glycol stearate, lithium stearate, magnesium stearate, manganese stearate, methyl stearate, potassium stearate, sodium stearate, strontium stearate, or zinc stearate, preferably aluminum stearate.

[0171] 18. The teat sealant composition of paragraph 17, wherein the composition comprises about 1% to 10% w / w of a metal salt of a long chain fatty acid.

[0172] 19. The teat sealant composition of paragraph 12 or paragraph 13, wherein the at least one thickening agent comprises silica, preferably hydrophilic or hydrophobic silica.

[0173] 20. The teat sealant composition of paragraph 19, wherein the composition comprises about 0.1% to 1% w / w silica, preferably hydrophilic fumed silica.

[0174] 21. The teat sealant composition of any of paragraphs 6 to 20, wherein:

[0175] - the solid particles are very soluble, readily soluble, soluble, slightly soluble, slightly soluble, very slightly soluble, practically insoluble or insoluble in water;

[0176] - the solubility of the solid particles in water exceeds about twice the isotonic concentration;

[0177] - the solid particles modify the rheology of the composition as well as provide antimicrobial activity, preferably against mastitis-causing bacteria;

[0178] - the solid particles comprise at least one type of salt other than a bismuth salt, such as an organic salt or an inorganic salt;

[0179] - the solid particles comprise a halide salt, a phosphate or a carbonate salt, preferably sodium chloride, potassium chloride, a calcium phosphate as defined herein (such as dicalcium phosphate dihydrate or dibasic calcium phosphate dihydrate) or magnesium carbonate;

[0180] - the solid particles contain at least one type of heavy metal salt, provided that it is not a bismuth salt, preferably the metal of the heavy metal salt contains calcium, magnesium or aluminum, but does not include toxic heavy metals;

[0181] - the solid particles comprise at least one type of carbohydrate, such as a simple sugar or a complex sugar, preferably sucrose or mannitol;

[0182] - the solid particles comprise at least one type of mineral, such as clay, such as kaolin, such as kaolinite, halloysite, dickite or nacrite;

[0183] - the solid particles contain sodium chloride;

[0184] - the solid particles contain potassium chloride;

[0185] - the solid particles comprise calcium phosphate as defined herein;

[0186] - the solid particles comprise dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate;

[0187] - the solid particles comprise calcium carbonate;

[0188] - the solid particles comprise magnesium carbonate;

[0189] - the solid particles comprise sodium chloride or potassium chloride and calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate);

[0190] - the solid particles contain sodium chloride or potassium chloride and calcium carbonate;

[0191] - the solid particles contain sodium chloride or potassium chloride and magnesium carbonate;

[0192] - the composition comprises solid particles in an approximate amount ranging from 10% to 75%, 35% to 70%, 40% to 65% or 50% to 60% w / w;

[0193] - the composition comprises 25-60% w / w sodium chloride or potassium chloride;

[0194] - the composition comprises 10-30% w / w magnesium carbonate;

[0195] - the composition comprises 10% w / w magnesium carbonate and approximately 25% w / w sodium chloride or potassium chloride;

[0196] - the composition comprises 55% w / w sodium chloride or potassium chloride;

[0197] - the composition comprises 35-60% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate);

[0198] - the composition comprises 60% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate);

[0199] - the composition comprises 10% w / w sodium chloride or potassium chloride and 50% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate);

[0200] - the composition comprises 25% w / w sodium chloride or potassium chloride and 35% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate); or

[0201] - The composition comprises 10% w / w sodium chloride or potassium chloride and 50% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate).

[0202] 22. The teat sealant composition of any of paragraphs 6 to 21, further comprising one or more other ingredients such as at least one antioxidant or preservative.

[0203] 23. A teat sealant composition comprising solid particles in the form of a gel or a paste or in the form of a coarse suspension formulated for occluding teat ducts and / or teat cistern.

[0204] 24. A teat sealant composition comprising:

[0205] an oily carrier; and

[0206] a thickener comprising solid particles and lauric acid,

[0207] wherein the solid particles are not bismuth salts.

[0208] 25. A teat sealant composition comprising a coarse filler material in the form of solid particles in a gelling agent, wherein the solid particles are not a bismuth salt.

[0209] 26. A teat sealant composition comprising:

[0210] Liquid paraffin;

[0211] Aluminum stearate;

[0212] fumed silica; and

[0213] Water-soluble solid particles,

[0214] The composition is in the form of a paste or a suspension.

[0215] 27. A teat sealant composition comprising:

[0216] Liquid paraffin;

[0217] Aluminum stearate;

[0218] fumed silica; and

[0219] Water-insoluble solid particles,

[0220] The composition is in the form of a paste or a suspension.

[0221] 28. The nipple sealant composition of any of paragraphs 23 to 27, wherein the average solid particle size is less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80 μm in size, or the average solid particle size is about 10-50 μm in size, or the average solid particle size is about 3-10 μm in size.

[0222] 29. The teat sealant composition of any of paragraphs 23 to 28, wherein:

[0223] - the solid particles are very soluble, readily soluble, soluble, slightly soluble, slightly soluble, very slightly soluble, practically insoluble or insoluble in water;

[0224] - the solubility of the solid particles in water exceeds about twice the isotonic concentration;

[0225] - the solid particles modify the rheology of the composition as well as provide antimicrobial activity, preferably against mastitis-causing bacteria;

[0226] - the solid particles comprise at least one type of salt other than a bismuth salt, such as an organic salt or an inorganic salt;

[0227] - the solid particles comprise a halide salt, a phosphate or a carbonate salt, preferably sodium chloride, potassium chloride, a calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate) or magnesium carbonate;

[0228] - the solid particles contain at least one type of heavy metal salt, provided that it is not a bismuth salt, preferably the metal of the heavy metal salt contains calcium, magnesium or aluminum, but does not include toxic heavy metals;

[0229] - the solid particles comprise at least one type of carbohydrate, such as a simple sugar or a complex sugar, preferably sucrose or mannitol;

[0230] - the solid particles comprise at least one type of mineral, such as clay, such as kaolin, such as kaolinite, halloysite, dickite or nacrite;

[0231] - the solid particles contain sodium chloride;

[0232] - the solid particles contain potassium chloride;

[0233] - the solid particles comprise calcium phosphate as defined herein;

[0234] - the solid particles comprise dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate;

[0235] - the solid particles comprise calcium carbonate;

[0236] - the solid particles comprise magnesium carbonate;

[0237] - the solid particles comprise sodium chloride or potassium chloride and calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate);

[0238] - the solid particles contain sodium chloride or potassium chloride and calcium carbonate;

[0239] - the solid particles contain sodium chloride or potassium chloride and magnesium carbonate;

[0240] - the composition comprises solid particles in an approximate amount ranging from 10% to 75%, 35% to 70%, 40% to 65% or 50% to 60% w / w;

[0241] - the composition comprises 25-60% w / w sodium chloride or potassium chloride;

[0242] - the composition comprises 10-30% w / w magnesium carbonate;

[0243] - the composition comprises 10% w / w magnesium carbonate and approximately 25% w / w sodium chloride or potassium chloride;

[0244] - the composition comprises 55% w / w sodium chloride or potassium chloride;

[0245] - the composition comprises 35-60% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate);

[0246] - the composition comprises 60% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate);

[0247] - the composition comprises 10% w / w sodium chloride or potassium chloride and 50% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate);

[0248] - the composition comprises 25% w / w sodium chloride or potassium chloride and 35% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate); or

[0249] - The composition comprises 10% w / w sodium chloride or potassium chloride and 50% w / w calcium phosphate as defined herein (such as dicalcium phosphate dihydrate / hydrogen calcium phosphate dihydrate).

[0250] 30. A teat sealant composition comprising one of the following formulations, wherein all ingredients shown are actual or approximate values:

[0251]

[0252] or

[0253]

[0254] or

[0255] NaCl or KCl 55% w / w;

[0256] Aluminum stearate 4.3% w / w;

[0257] Fumed silica 0.7% w / w;

[0258] Lauric acid 2.5% w / w; and

[0259] Paraffin oil 37.5% w / w;

[0260] or

[0261] NaCl or KCl 10% w / w;

[0262] Calcium phosphate as defined herein 50% w / w; Aluminium stearate 4.3% w / w;

[0263] Fumed silica 0.7% w / w;

[0264] Lauric acid 5% w / w; and

[0265] Paraffin oil 30% w / w;

[0266] or

[0267] NaCl or KCl 25% w / w;

[0268] Calcium phosphate as defined herein 35% w / w; Aluminium stearate 4.3% w / w;

[0269] Fumed silica 0.7% w / w;

[0270] Lauric acid 5% w / w; and

[0271] Paraffin oil 30% w / w;

[0272] or

[0273] Calcium phosphate as defined herein 60% w / w; Aluminium stearate 4.3% w / w;

[0274] Fumed silica 0.7% w / w;

[0275] Lauric acid 5% w / w; and

[0276] Paraffin oil 30% w / w;

[0277] or

[0278] Magnesium carbonate 25% w / w;

[0279] Aluminum stearate 4.3% w / w;

[0280] Fumed silica 0.7% w / w;

[0281] Lauric acid 5% w / w; and

[0282] Paraffin oil 65% w / w;

[0283] or

[0284] NaCl or KCl 50% w / w;

[0285] Aluminum stearate 5.3% w / w;

[0286] Fumed silica 0.7% w / w;

[0287] Lauric acid 5% w / w; and

[0288] Paraffin oil 39% w / w;

[0289] or

[0290] Magnesium carbonate 30% w / w;

[0291] Fumed silica 0.7% w / w;

[0292] Lauric acid 5% w / w; and

[0293] Paraffin oil 64.3% w / w;

[0294] or

[0295] NaCl or KCl 50% w / w;

[0296] Aluminum stearate 5.3% w / w;

[0297] fumed silica 0.7% w / w; and

[0298] Paraffin oil 44% w / w;

[0299] or

[0300] NaCl or KCl 25% w / w;

[0301] Magnesium carbonate 10% w / w;

[0302] Aluminum stearate 4.3% w / w;

[0303] fumed silica 0.7% w / w; and

[0304] Paraffin oil 60% w / w.

[0305] 31. A method of preparing a teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle, the method comprising the steps of:

[0306] (1) forming a water-insoluble gel by mixing at least one type of water-insoluble carrier agent with at least one type of thickening agent;

[0307] (2) adding solid particles to the gel to form a paste; and, optionally,

[0308] (3) adding at least one other type of thickening agent to the gel or paste, wherein:

[0309] preparing a teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle;

[0310] The solid particles are not bismuth metal salts;

[0311] Step (3) is optional; and

[0312] Steps (2) and (3) do not need to be performed in the order described.

[0313] 32. A method of preparing a teat sealant composition, the method comprising the steps of:

[0314] (1) forming a water-insoluble gel by mixing at least one type of oily vehicle with at least one type of thickening agent;

[0315] (2) adding solid particles to the gel to form a paste; and,

[0316] (3) adding lauric acid as a thickening agent to the gel or paste, wherein:

[0317] The solid particles are not bismuth metal salts.

[0318] 33. A teat sealant composition prepared by the method according to paragraph 31 or paragraph 32.

[0319] 34. A teat sealant composition for preventing new intramammary infection or mastitis in a cow, wherein the composition is as described in any one of paragraphs 1 to 30 and 33.

[0320] 35. A teat sealant composition for preventing new intramammary infection or mastitis in a cow or when used for such use, wherein the composition is as described in any one of paragraphs 1 to 30 and 33.

[0321] 36. Use of a teat sealant composition in the preparation of a medicament for preventing new intramammary infection or mastitis in a cow, wherein the composition is as described in any one of paragraphs 1 to 30 and 33.

[0322] 37. A method of preventing new intramammary infection or mastitis in a cow, the method comprising the step of applying the composition described in any one of paragraphs 1 to 30 and 33 to at least one teat of the cow.

[0323] 38. Use of a composition as described in any one of paragraphs 1 to 30 and 33 for preventing new intramammary infection or mastitis in a cow.

[0324] 39. A syringe containing the composition described in any one of paragraphs 1 to 30 and 33.

[0325] 40. A kit for use in a method of preventing a new intramammary infection or mastitis in a cow, or when used for such use, wherein the kit comprises: a syringe capable of administering the composition described in any one of paragraphs 1 to 30 and 33 to at least one teat of the cow.

[0326] 41. The teat sealant composition of paragraph 34 or 35, the use of paragraph 36 or 38, the method of paragraph 37, or the kit of paragraph 40, wherein the composition is used in a dairy cow or heifer.

[0327] 42. The teat sealant composition of paragraph 34, 35, or 41, the use of paragraph 36, 38, or 41, the method of paragraph 37 or 41, or the kit of paragraph 40 or 41, wherein:

[0328] - the composition prevents new intramammary infection or mastitis for a period of at least about 14 days, about 42 to about 90 days, or up to about 4 months;

[0329] The blocking effect of the composition is for the first 14 days of the dry period, until the natural keratin plug is formed;

[0330] - the composition occludes the teat duct and / or teat cistern or substantially occludes the teat duct and / or teat cistern until a natural keratin plug has formed in the teat duct;

[0331] - the composition protects the cow's teat(s) from bacterial infection throughout the dry period;

[0332] - administering the composition to each quadrant at the end of lactation;

[0333] - administering the composition per quadrant in primiparous heifers approximately 4 weeks prior to calving;

[0334] - the teat sealant composition has a sufficiently low viscosity to allow it to be administered by injection through the teat canal and / or teat cistern;

[0335] - Apply / inject approximately 2-5g of the teat sealant composition into the teat canal and / or teat cistern;

[0336] - the injection force for injecting the teat sealant composition into the teat at 25°C does not exceed 4500g, and preferably the injection force for injecting the teat sealant composition into the teat at 25°C is between about 1500g and 3500g;

[0337] - the teat sealant composition rapidly thickens in the teat, thereby being retained and blocking the teat ducts and / or teat cistern;

[0338] - once located within the teat canal and / or teat cistern, the teat sealant composition has a sufficiently high viscosity to be retained within and block the teat canal or teat cistern;

[0339] - the teat sealant composition has a rebound viscosity enabling it to be retained within the teat canal and / or teat cistern as a substantially cohesive mass;

[0340] The teat sealant composition has rheological properties that enable the composition to be administered into the teat duct and / or teat cistern by injection and to be retained in the teat duct and / or teat cistern to block the teat duct and / or teat cistern or to prevent intramammary infection;

[0341] - the teat sealant composition shear thins under the shear forces applied during injection and expression from the teat; or

[0342] - The teat sealant composition occludes or substantially occludes the teat ducts and / or teat cistern for about 4-6 weeks while allowing the natural keratin plug to form.

[0343] Features described with respect to a composition / formulation / medicament may also relate to a method / use, and vice versa, if the context permits. It will be understood that the terms "formulation", "medicament" and "composition" may be used interchangeably, if the context permits.

[0344] Having broadly described the present invention in its various embodiments, non-limiting examples of preferred embodiments will now be described.

[0345] BRIEF DESCRIPTION OF THE DRAWINGS

[0346] Figure 1 of Example 1

[0347] Figure 1 .Symable properties of F1 after storage at 4, 25 and 40°C for different periods of time.

[0348] Figure 2 .Symable properties of F2 after storage at 25 and 40°C for different periods of time.

[0349] Figure 3 .Teatseal TM and flow behavior of calcium carbonate and calcium phosphate preparations.

[0350] Figure 4 .Teatseal TM and injection simulations of calcium carbonate and calcium phosphate preparations.

[0351] Figure 5 .Teatseal TM and the flow behavior of calcium phosphate formulations containing higher aluminum stearate and 5% lauric acid.

[0352] Figure 6 .Teatseal TM and injection simulations of calcium phosphate formulations containing higher aluminum stearate and 5% lauric acid.

[0353] Figure 7 .Teatseal TMand flow behavior of prototype formulations.

[0354] Figure 8 .Teatseal TM and injection simulations of prototype formulations.

[0355] Figure 9 . Recovery (%) of the formulations tested in the suspended udder study of Example 7.

[0356] Figure 10 Effect of storage at 25°C on syringeability (syringability).

[0357] Figure 11 Effect of storage at 25°C on syringeability (syringability).

[0358] Figure 12 Effect of temperature cycling (4 and 25°C) on the injectability of calcium phosphate and sodium chloride formulations.

[0359] Figure 13 Storage stability of calcium phosphate and sodium chloride preparations at 25°C.

[0360] Figure 14 .Flow behavior of NaCl and KCl preparations.

[0361] Figure 15 .Injection simulation behavior of NaCl and KCl preparations.

[0362] Figure 2 of Example 2

[0363] Figure 16 .Commercially available Teatseal TM flow behavior.

[0364] Figure 17 .Teatseal TM Injection simulation behavior of the batch.

[0365] Figure 18 .Teatseal TM Injectability of the batch.

[0366] Figure 3 of Example 3

[0367] Figure 19 Effect of the method of addition of lauric acid on the rheological properties (viscosity versus shear rate) of the formulation: light green - bottom line of the graph (method 1), dark green - top line of the graph (method 2).

[0368] Figure 20 .Effect of lauric acid addition method on rheological properties during injection simulation test.

[0369] Figure 21 Effect of adding lauric acid to room temperature formulations versus warmed formulations on formulation rheology (viscosity versus shear rate): blue (room temperature formulation), orange (warmed formulation).

[0370] Figure 22 Effect of adding lauric acid to room temperature formulations versus warmed formulations on rheological properties during injection simulations.

[0371] Figure 4

[0372] Figure 23 .With Teastseal TM and Biobloc TM Comparison of the flow behavior of various experimental teat sealant formulations.

[0373] Figure 24 .With Teastseal TM and Biobloc TM Compared to injection simulations of various experimental teat sealant formulations.

[0374] Figure 25 .With Teastseal TM and Biobloc TM Comparison of the flow behavior of various experimental teat sealant formulations.

[0375] Figure 26 .With Teastseal TM and Biobloc TM Compared to injection simulations of various experimental teat sealant formulations.

[0376] Figure 27 .With Biobloc TM Compared to the experimental teat sealant formulations the injectability was simulated.

[0377] Figure 28 .With Teastseal TM and Biobloc TM Comparison of the flow behavior of various experimental teat sealant formulations containing calcium carbonate.

[0378] Figure 29 .With Teastseal TM and Biobloc TM Compared to injection simulations of various experimental teat sealant formulations containing calcium carbonate.

[0379] Figure 30 .With Biobloc TM Comparison of the injectability of various experimental teat sealant formulations containing calcium carbonate.

[0380] Figure 31 .With Teastseal TM and Biobloc TM The flow behavior of various experimental teat sealant formulations containing dibasic calcium carbonate was compared.

[0381] Figure 32 .With Teastseal TM and Biobloc TM Compared to injection simulations of various experimental teat sealant formulations containing dibasic calcium carbonate.

[0382] Figure 33 The experimental teat sealant formulations delivered from either the BioBloc syringe or the white HDB syringe were compared to the BioBloc TM Compared to injectable.

[0383] Figure 34 Biobloc, which uses its own syringe TM Injectability.

[0384] Figure 35 Biobloc using a white HDB syringe TM Injectability.

[0385] Figure 36 Injectability of the trial teat sealant formulation delivered using a white HDB syringe.

[0386] Figure 37 Injectability of the experimental teat sealant formulation delivered using a Biobloc syringe.

[0387] Figure 38 Photographs of the behavior of a test teat sealant formulation in a water-filled syringe simulating a teat.

[0388] Figure 39 Photographs of the behavior of experimental teat sealant formulations in water simulating milk.

[0389] Figure 5 of Example 5

[0390] Figure 40. Solution killing of E. coli and S. uberis by various paste formulations was evaluated and presented as a graph of bacterial cell survival. A. Solution killing of E. coli using various pastes within 24 hours. B. Solution killing of E. coli using various pastes within 48 hours. C. Solution killing of S. uberis using various pastes within 48 hours.

[0391] Figure 41. Counts of Streptococcus uberis cells exposed to teat sealant formulations over time. A. Streptococcus uberis treated with a control formulation (prototype) over 5 days. B. Streptococcus uberis treated with a 20% NaCl formulation (prototype) over 5 days. C. Streptococcus uberis treated with a 30% NaCl formulation (prototype) over 5 days. D. Streptococcus uberis treated with a 40% NaCl formulation (prototype) over 5 days. E. Streptococcus uberis treated with a 55% NaCl formulation (prototype) over 5 days. F. Streptococcus uberis treated with various formulations (prototype) over 5 days. G. Streptococcus uberis treated with a KCl formulation (prototype) over 5 days. H. Streptococcus uberis treated with a kaolin formulation (prototype) over 5 days.

[0392] Figure 42. E. coli cell counts over time after exposure to different teat sealant formulations. A. Average survival curves of E. coli treated with different teat sealant formulations over 20 days. B. E. coli treated with different teat sealant formulations (such as KCl) over 5 days.

[0393] Figure 43. Staphylococcus aureus cell counts over time following exposure to teat sealant paste formulations. A. Survival curves of Staphylococcus aureus treated with different teat sealant prototype formulations over 30 days. B. Average survival curves of Staphylococcus aureus treated with different teat sealant prototype formulations over 20 days.

[0394] Figure 6 of Example 6

[0395] Figure 44 Ultrasound image of the left front teat of cow 302, 1 hour (F2).

[0396] Figure 45 Ultrasound image of the right front teat of cow 302, 1 hour (F3).

[0397] Figure 46 Ultrasound image of the right hind teat of cow 302, 1 hour (Teatseal TM ).

[0398] Figure 47 Ultrasound image of the left hind teat of cow 302, 1 hour (F1).

[0399] Figure 48 Mammary glands of cows 28 days after treatment (dry period) show some milk present in the udder but not yet completely involuted (top row) and some udders that have completely involuted (bottom row).

[0400] Figure 49 Schematic representation of teat sealant scoring. The red shaded area indicates teat sealants.

[0401] Figure 50 .1 hour sample; % recovery relative to the injected formulation dose.

[0402] Figure 51 .48 hour sample; % recovery relative to the injected formulation dose.

[0403] Figure 52 .672 hour sample; % recovery relative to injected formulation dose.

[0404] Figure 53 .672 hour sample; % recovery relative to injected formulation dose.

[0405] Figure 54 .1 hour sample; % recovery relative to TS.

[0406] Figure 55 .48 hour sample; % recovery relative to TS.

[0407] Figure 56 .48 hour sample; % recovery relative to TS.

[0408] Figure 57 .672 hour sample; % recovery relative to TS.

[0409] Figure 58 .672 hour sample; % recovery relative to TS.

[0410] Figure 7

[0411] Figure 59 Papillary lengths were grouped according to the treatment assigned to each papilla. Bars represent median values, which are also depicted at the bottom of each bar. Error bars represent ranges, and points represent individual points (n=12 / treatment).

[0412] Figure 60 The percentage of solids in milk samples was grouped according to the treatment assigned to each teat. The bars represent the median value, which is also depicted at the bottom of each bar. The error bars represent the range, and the points represent individual points (n=12 / treatment).

[0413] Figure 61 The mass of the formulation injected into the nipple was grouped by treatment. The bars represent the median value, which is also depicted at the bottom of each bar. The error bars represent the range, and the points represent individual points (n=12 / treatment).

[0414] Figure 62Estimated recovery by dripping (A), squeezing (B), scraping (C), and flushing (D), as well as the total recovery of the infusion formulation (E). The bars represent the median, which is also depicted at the bottom of each bar. Error bars represent the range, and points represent individual points (n=12 / treatment).

[0415] Figure 63 Estimated recovery from nipples. Bars represent median values, which are also depicted at the bottom of each bar. Error bars represent ranges, and points represent individual points (n=12 / treatment).

[0416] Figure 64 Scatter plot of nipple length and percent recovery from the nipple with linear regression line (solid line) and 95% confidence interval (dashed line).

[0417] Figure 65 The recovery rate of the front teat (A) and the back teat (B) is greater than or equal to that of Teatseal TM The recovery rate of the anterograde enzyme was ≥95.81% in the anterior teat and ≥84.64% in the posterior teat.

[0418] Figure 66 .The incised papilla is shown with the remaining preparation after expression and before scraping.

[0419] Figure 8

[0420] Figure 67 .Location of breast samples for histopathology.

[0421] DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0422] Example 1 - Development and Use of Teat Sealant Formulations

[0423] background

[0424] In dairy cows, the incidence of mastitis caused by environmental pathogens is highest at the beginning of the 7-10 week dry period and again at calving at the end of the dry period. During the dry period, cows develop a natural plug, the keratin plug, which seals the teat canal; however, this takes some time (several weeks) during which microorganisms can enter, necessitating the use of internal teat sealants.

[0425] Teat sealants are used in the dry period of dairy cows and heifers to "seal" the teat canals and thereby prevent, or at least reduce, the entry of environmental pathogens into the udder.

[0426] The formulation of teat sealants presents the following challenges:

[0427] - It must be fluid enough so that 2-5 g can be injected into a single teat duct via a syringe (without a needle).

[0428] -It should flow into the epithelial folds of the mucosal lining of the nipple to seal most effectively.

[0429] - It must then thicken so that it remains in the teat cistern and possibly in the teat (striate) ducts during the dry period (up to 10 weeks).

[0430] - It must be able to be squeezed out at the end of the dry period.

[0431] - It preferably remains as a cohesive mass in the teat; that is, some of the preparation should not break off and enter the udder and thus contaminate the first milking after the dry period.

[0432] The ideal rheological properties of teat sealant formulations are unclear. Furthermore, it is unclear whether the sealant should remain in the teat throughout the dry period, or whether it only needs to be present for the first 4-6 weeks, while the natural keratin plug is allowed to form. This has the advantages of not requiring milking at the end of the dry period and being less likely to be contaminated with teat sealant during the first milking.

[0433] Most teat sealant preparations use high concentrations of bismuth subnitrate suspended in liquid paraffin. These have the following disadvantages:

[0434] - The first milking after the contaminated dry period.

[0435] - Clogged filter in milking line.

[0436] - Downstream processing problems, such as block-spot in cheese.

[0437] The goal of this study was to replace teat sealants containing the heavy metal bismuth subnitrate with water-soluble or insoluble, environmentally acceptable materials. All formulations were high-concentration suspensions of these materials in an acceptable vehicle. The theory of such high-concentration suspensions has generated interest in certain non-pharmaceutical sectors (e.g., concrete manufacturers, oil drillers), but predictive theory is poor, making empirical studies crucial.

[0438] Without wishing to be bound by theory, it appears that the rheological properties of concentrated suspensions are influenced by the following factors:

[0439] -Solids concentration.

[0440] - Particle size, size distribution, particle shape, particle surface properties, adhesion properties.

[0441] - Rheological properties of the suspension (Newtonian, non-Newtonian).

[0442] -History: preparation method, storage conditions and time.

[0443] Teatseal TM It is a commercial product known for its high efficiency. The inventors' purpose is to understand the TM The rheological properties of the bismuth subnitrate were determined (see Example 2) and this information was then used to guide the development of formulations using soluble and insoluble solid particles / fillers in place of bismuth subnitrate.

[0444] Initial studies attempted to create a suitable viscoelastic system using ethylcellulose, castor wax, chitosan, and alginate zein to match the Teatseal TM performance, but this was unsuccessful. The decision was then made to use a highly concentrated oil suspension rather than attempting a gelling polymer system. This was a phased, iterative process in which alternative materials to bismuth subnitrate were gradually screened and either eliminated based on their performance in screening trials or selected for further trials. TM According to the evaluation of (see Example 2), aluminum stearate was retained as a gelling agent for the oil.

[0445] Initially, Eudragit RLPO microparticles were tried as solid particles / fillers but were abandoned because it was a non-biodegradable polymer, which made it environmentally unacceptable. The inventors then turned to water-insoluble environmentally acceptable excipients such as magnesium carbonate, kaolin, microcrystalline cellulose (Avicel TM ) and keratin.

[0446] The volume fraction of suspended solids was considered important when attempting to understand the rheological properties of concentrated oil suspensions. To determine the volume fraction of solids, the inventors needed to use solids that did not absorb oil, as this would reduce the "free" oil and thus alter the solid volume fraction calculation. Therefore, the inventors decided to use sodium chloride as one of the solids. This led to the serendipitous discovery of the possibility of using water-soluble fillers / solid particles to create teat sealants.

[0447] The idea of ​​using soluble fillers / particles is clearly novel and opens up the possibility of designing slowly eroding teat sealants, thus overcoming the need for milking out at the end of the dry period.

[0448] Teatseal TM The reasons for the effectiveness of are unclear. It is thought that, at least in part, it acts as a physical barrier to prevent bacteria from entering the nipple. It may also be due to its antibacterial properties. Therefore, another aspect of formulation development is testing the antibacterial properties of the developed formulation.

[0449] Materials / Excipients

[0450] Paraffin oil-BP grade, lauric acid, magnesium carbonate, bismuth subnitrate (Sigma-Aldrich), aluminum stearate (Sterm Chemicals, USA), Aerosil (Aerosil-200 and Aerosil R972), Teatsea TM (Zoetis), NaCl (Dominion Salt Limited), Avicel PH 112 (FMC corporation) and kaolin (heavy) (Kempthorne Prosser & Co Limited) were obtained from commercial sources. NaCl was ground in a planetary ball mill (Retsch PM-100, ball size 9 mm) for 1 hour (speed - 600 rpm) and sieved for 30 minutes (90 μm).

[0451] Preparation method for laboratory testing (beaker method)

[0452] Weighed amounts of paraffin oil and aluminum stearate were added to a beaker and heated at 120°C for 30 minutes. After the mixture was melted / heated, the beaker was removed from the heat source and mixed with a spatula until a gel-like mixture formed. The gel was allowed to cool at room temperature, followed by the addition of solid particles / fillers and thorough mixing with a spatula. After the fillers were mixed, Aerosil-200 was added and mixed thoroughly with a spatula. Lauric acid was melted at 45°C and added to the mixture, mixing thoroughly with a spatula. The formulation was allowed to stand in the beaker at room temperature for 24 hours before testing.

[0453] Preparation Methods for Retention Studies

[0454] Syringes, aluminum stearate, Aerosil-200, Aerosil-R972, lauric acid, kaolin, and paraffin oil were sterilized by gamma irradiation. Magnesium carbonate, sodium chloride, and glassware were sterilized using heat sterilization (160°C, 3 h). Each batch of formulation was prepared using the beaker method in 100 g portions.

[0455] Injectability

[0456] Unless otherwise noted, a standard 4.5 ml (or larger) intramammary syringe sold by Hubert De Backer nv (HDB Veterinary Intramammary Syringe) with a collar / nozzle inner diameter of 1.5 mm and an inner barrel diameter of 13.5 mm (https: / / www.hdb.be / _library / _files / Tabellen / 0.5.0 / Intramammary_0_5_0.pdf) was used in each of the examples described herein.

[0457] A texture analyzer was used to measure the force and work required to expel a formulation from a syringe. The syringe plunger was advanced at a constant speed of 2 mm / s over a distance of 25 mm to expel the formulation in 12.5 seconds. The analyzer continuously measured force, providing a graph of force versus time.

[0458] Rheological measurements

[0459] The rheological properties of the formulations were tested using a Discovery Hybrid Rheometer HR-3 (TAinstruments). The sample (4-5 g) was placed on the plate of the rheometer. The cone / plate geometry was: 2°, 60 mm, and a gap of 500 μm. The temperature was fixed at 32°C.

[0460] Shear thinning was tested by applying a shear of 0 to 100 1 / s. The soak time and hold time were 300 s. To test the rebound performance, i.e. the rate of recovery of viscosity after shear thinning, an initial shear rate of 550 1 / s was applied, followed by shear rates of 10 and 1 1 / s.

[0461] Centrifugation / Phase Separation

[0462] 10 g of the preparation was added to a 15 mL Falcon tube and centrifuged for 30 minutes at 1800 G. After centrifugation, the phase separation (oil layer) was visually assessed.

[0463] Dispersion / swelling

[0464] To test dispersion, each formulation (accurately weighed 3-4 g) was added to a Falcon tube (50 mL) followed by 40 mL of distilled water. In another experiment, the formulation was added to a scintillation vial and 20 mL of distilled water was added. The vial and test tube were shaken at 37° C. on a rotary shaker (150 rpm). In another experiment, 20 mL of milk was added instead of water.

[0465] The weight of the remaining formulation was determined at various times by decanting the water, drying the vial containing the remaining formulation at 60°C for 24 hours, and weighing. In this case, the vial contained milk, and after decanting the milk, the remaining formulation was gently rinsed three times with 10 mL of water to remove milk solids and then dried as above.

[0466] In another screening test for swelling and dispersion, the formulation (3-4 g) was added to a Falcon tube and then water was added. The swelling of the formulation was assessed after 12-24 hours and then gently shaken. Some non-dispersed formulations were stored for a long time in a Falcon tube with excess water to determine the long-term consistency of the formulation in an aqueous environment.

[0467] To increase understanding of the dispersion of the formulation in an aqueous medium, the formulation (0.5 g) was added to a 15 mL Falcon tube and 10 mL milliQ water was added. The tube was shaken at 150 rpm for 5 days in a 37° C. incubator, then centrifuged at 3000 rpm for 30 minutes, and the supernatant was removed. The supernatant was passed through a 0.22 μm filter and analyzed using inductively coupled plasma mass spectrometry (ICPMS). MilliQ water was used as a control and analyzed for sodium, bismuth, and magnesium ions.

[0468] Retention Research

[0469] The formulation was administered to groups of cows according to a balanced design as described in the Examples below. One hour, two days, and two or four weeks after administration, teats were milked to recover the remaining teat sealant formulation. The recovered material was quantified as follows.

[0470] Receive frozen sample tubes (Falcon) and store at -18°C. For analysis, the Falcon tubes were taken out of the refrigerator and equilibrated overnight at room temperature. Record the weight (wet weight) of the tubes containing the samples. Record the weight of the empty culture dishes and pour the samples into the culture dishes. The samples in the tubes and culture dishes were dried at 50°C for 24 hours. After drying, record the weight of the tubes and culture dishes. After deducting the empty weights of the tubes and culture dishes, calculate the total dry weight of the sample. Use the following equation to deduct the milk solids in the dry sample and calculate the weight of the preparation recovered.

[0471] Weight of milk solids in dry sample = (total weight of wet sample - total weight of dry sample) / 0.9 x 0.1

[0472] Formulation weight: total weight of dry sample - calculated milk solids weight.

[0473] Note: The mass loss (g) during the 24 h drying period at 50°C is attributed to the moisture in the sample milk, assuming that the milk contains 10% solids and 90% water. It is also assumed that there are no volatile components in the formulation that are lost during heating to 50°C for 24 hours.

[0474] Iteration 1: Laboratory trials of retention studies

[0475] Material / filler screening and selection

[0476] In these initial screening studies, formulations were prepared using the beaker method.

[0477] Table 1A. Initial screening of fillers / solid particles for teat sealant formulations.

[0478]

[0479]

[0480] All formulations contained paraffin oil (to balance). Laboratory grade MgCO3 was used.

[0481] The particle size of commercial sodium chloride is too large to make a smooth formulation. It was ground using a planetary ball mill (Retsch PM-100) for 1 hour and then sieved to break up agglomerates. The ground and sieved NaCl was stored in an airtight container and used for the NaCl formulation.

[0482] Dispersion / swelling behavior of the formulation in water

[0483] The keratin formulation was allowed to swell and disperse in water. After a week, the Avicel formulation was allowed to swell in water. Keratin and Avicel were ultimately deemed unsuitable.

[0484] The magnesium carbonate (35%), kaolin (45%) and NaCl (50%) formulations did not swell in water. The NaCl formulation was found to be intact and surprisingly, the sodium chloride did not leach out under the experimental conditions.

[0485] Parallel microbiological tests (see Example 5) showed that it was necessary to add antimicrobial agents to certain formulations to achieve teatseal TM Therefore, lauric acid (5%, 10%) is included.

[0486] ICPMS analysis

[0487] ICPMS tests were performed to check the dissolution of fillers / solid particles in water. From Teatseal TM The levels of bismuth ions and magnesium are related to Na + The ion ratio is low, indicating that the sodium chloride formulation has dissolved. The dissolution of NaCl depends on the percentage of aluminum stearate, indicating that it is possible to adjust the dispersion of the sodium chloride formulation by changing the percentage of aluminum stearate. This study used water (Milli Q) as a control and showed that magnesium and sodium ions are present in it, which is why Teatseal TM The samples also showed low concentrations of these ions.

[0488] Table 1B. ICPMS analysis of water from dispersion experiments.

[0489]

[0490]

[0491] All formulations contained 5% LA and 0.7% Aerosil.

[0492] Formulation for retention studies in cows (see Example 6)

[0493] Following this screening work, magnesium carbonate, sodium chloride, and kaolin were selected as fillers / solid particles in formulations for evaluation in cows. Six formulations were selected and screened based on their dispersibility, rheology, and syringeability. Preliminary storage stability was assessed in conjunction with retention studies.

[0494] Table 1C. Concentrations (% w / w) of ingredients in the formulations used for the retention studies of Example 6.

[0495] Element F1 F2 F3 F4 F5 F6 <![CDATA[MgCO3]]> 25％ 30％ 10％ NaCl 50％ 50％ 25％ Kaolin 45％ Aluminum stearate 4.3％ 5.3％ 4.3％ 5.3％ 4.3％ Aerosil 0.7％ 0.7％ 0.7％ 0.7％ 0.7％ 0.7％ Lauric acid 5％ 5％ 5％ 5％

[0496] F4 contained Aerosil-R972, and the other formulations contained Aerosil-200 ('Aerosil').

[0497] Weight loss of formulation in milk or water (gravimetric analysis)

[0498] Milk has a slightly acidic pH (pH 6.6), so prototype formulations were tested to determine dispersion / dissolution in this medium. Specifically, the inventors were interested in testing the MgCO3 formulation with milk. The inventors tested the MgCO3 and NaCl formulations for four weeks. The NaCl formulation only lost weight when milk was added, while the MgCO3 formulation did not lose weight when milk or water was added.

[0499] Table 1D. NaCl loss in F2 formulation.

[0500]

[0501]

[0502] Table 1E. MgCO3 loss in F1 formulation.

[0503]

[0504] Rheology and syringeability

[0505] The goal is to make the injection force lower than 4500g. All preparations meet this standard. Two magnesium carbonate preparations F1 and F3 were selected because F1 shows good rheology and injectability, and F3 was selected as an example of a poor preparation because of its viscosity and especially its poor rebound viscosity. Similarly, two preparations (F2 and F5) containing the water-soluble salt NaCl were selected. F2 contains lauric acid, and F5 does not contain lauric acid. The inventor's earlier work found that the kaolin preparation made with AerosilR972 has better dispersibility (no swelling) in water than the preparation made with Aerosol-200. The F6 preparation has used a combination of NaCl and MgCO3.

[0506] Table 1F. Retention of rheological properties and syringeability of the study formulations.

[0507]

[0508] F4 contained Aerosil-R972 and the other formulations contained Aerosil-200. ND = Not Determined.

[0509] Centrifugation (phase separation)

[0510] As the inventor noticed that Teatseal TM Phase separation during storage occurs, so this test is performed as a stress test. TM The prototype formulations were centrifuged (18000 G force) for 30 minutes and then visually inspected. TM Oil separation occurred in this stress test, which did not occur with the prototype formulation.

[0511] Storage stability

[0512] Formulation F1 was stored at 4, 25, and 40°C for five weeks and the injectability test was performed at different times during this period. F1 showed good stability at 4°C (force below 4000 g), but at 25 and 40°C, hardening of the formulation was observed over time. See Example 1- Figure 1 It was therefore decided to discontinue the use of magnesium carbonate (although it continues to be tested in the current retention study).

[0513] Formulation F2 was stored at 25 and 40°C for four weeks. The sodium chloride formulation remained physically stable and hardened during storage. Figure 2 .

[0514] Retention Research

[0515] Details of methodology and analysis can be found in Example 6.

[0516] A retention study was conducted in dairy cows. Six prototype formulations and Teatseal were tested in this trial. TM The formulations were tested in four treatment groups: see Example 6 for a detailed discussion of the results.

[0517] Table 1G. Treatment summary table.

[0518] Treatment group preparation Time (h) Group 1 F1-F3+TS 1 Group 2 F1-F3+TS 48 Group 3 F1-F3+TS 672 Group 4 F4-F6+TS 672

[0519] TS=Teatseal TM

[0520] Table 1H. Recovery rate (%) of the formulation at different times after administration.

[0521]

[0522] The data vary widely. To test the method, a recovery at 1 hour was performed. A low recovery at 1 hour could indicate:

[0523] - The preparation is not properly squeezed out of the nipple.

[0524] - Systematic errors in the quantification method (e.g. over-deduction of milk solids).

[0525] Low recoveries at 1 hour may also be due to:

[0526] - During the first hour, the preparation drips from the nipple.

[0527] - Some are applied into the breast (although the nipple should be clamped at the base during application).

[0528] These recoveries at 1 hour were lower than those found in several previous retention studies. The low recoveries in some cows at 1 hour mean that caution should be exercised in interpreting the data from this trial, but the following observations were made:

[0529] - F1 vs. F3: F3 is said to be a poor formulation (no aluminum stearate, poor rebound), so perhaps the requirement for high rebound could be relaxed.

[0530] - F1, F3 vs. F2, F5: Formulations containing soluble sodium chloride (F2, F5) were retained in some cows for 672 hours, as were those containing insoluble magnesium carbonate (F1, F3).

[0531] Iteration 2: Laboratory Trials of the Suspended Breast Study

[0532] In this iteration, additional solid particles / fillers were investigated and the impact of the manufacturing method was studied. A hanging breast trial was conducted but development was terminated before the planned retention study could be conducted.

[0533] Preparation method

[0534] Beaker method

[0535] As above

[0536] Scraper method

[0537] Preliminary experiments showed that the preparation method affects the rheological properties of the formulation. The temperature, time and order of adding ingredients must be standardized.

[0538] Aluminum stearate and paraffin oil were added to a beaker and heated in an oven preheated to 120°C for 40 minutes to melt the aluminum stearate. The beaker was removed from the oven and the mixture was thoroughly mixed with a spatula to form a gel. The gel was cooled to room temperature. The gel and Aerosil-200 were mixed on a glass plate with a spatula. The filler (solid particles) was then added and thoroughly mixed with a spatula and stirred until a smooth formulation was obtained. The formulation was placed in a beaker and melted lauric acid (45°C) was added. The mixture was poured back onto the plate and any lauric acid lumps were broken up with a spatula.

[0539] Suspended breast preparation

[0540] The formulations were prepared by the spatula method. A hanging breast formulation was prepared using 0.3% methylene blue dye. The dye was added to the aluminum stearate and oleogel, stirred with a spatula, and then Aerosil-200 was added. These formulations were prepared using the spatula method.

[0541] Preparation of sterile preparations for antimicrobial testing

[0542] Sterilize the laminar flow hood. Wrap all clean glassware and spatulas with aluminum foil and perform heat sterilization (160°C, 3 hours). Add sodium chloride to a glass beaker and perform heat sterilization. Since calcium phosphate (dicalcium phosphate dihydrate, which is calcium hydrogen phosphate dihydrate) cannot be heat sterilized, a thin layer of calcium phosphate powder is sterilized with ultraviolet radiation for 30 minutes in a laminar flow hood. All preparations are made using the spatula method. After manufacturing the preparation, a thin layer of the preparation is spread on a glass plate and sterilized with ultraviolet radiation for 30 minutes in a laminar flow hood. After final sterilization, the preparation is packaged in a sterile Falcon tube.

[0543] Analysis of samples from the hanging breast study

[0544] The hanging udder samples were frozen upon receipt and stored at -18°C until analysis. After thawing overnight, the tubes were wiped with a paper towel and the wet weight of the capped tubes was recorded. The tubes were dried in an oven preheated to 50°C for 24 hours. After drying, the weight of the capped tubes was recorded.

[0545] The study was terminated early and these breast samples were used for subsequent studies as described in Example 7.

[0546] Density of fillers / solid particles

[0547] Since the density of the formulation may affect retention at the nipple base, this should be considered when selecting alternative materials.

[0548] Table II: Reported filler density.

[0549] Fillers <![CDATA[Density (g / cm 3 )]]> bismuth subnitrate 4.93 magnesium carbonate 2.16 Sodium chloride 2.16 calcium carbonate 2.71 Calcium phosphate (dicalcium phosphate dihydrate) 2.93

[0550] Calculated formulation density

[0551] The density of the formulation is calculated by taking into account the mass of the filler and the oil. For example, Teatseal TM Contains 65% w / w bismuth subnitrate and 35% w / w paraffin oil. The density of the prototype formulation was calculated using the following method:

[0552] Density of bismuth subnitrate = 4.93 g / cm 3

[0553] Density of paraffin oil = 0.9 g / cm 3

[0554] Density (ρ) = mass (m) / volume (V)

[0555] Volume of bismuth subnitrate = 65 / 4.93 = 13.18

[0556] Paraffin oil volume = 35 / 0.9 = 38.8

[0557] Volume (total) = 51.98 - Density assuming volume is additive = 100 / 51.98 = 1.92 g / cm 3

[0558] Table 1J. Calculated formulation densities.

[0559]

[0560]

[0561] Note: Microbiological testing has shown that it is necessary to add antimicrobial agents to certain formulations to achieve Teatseal TM Therefore, lauric acid (5%, 10%) is included.

[0562] Based on these density calculations, calcium carbonate and calcium phosphate were included in the screening program.

[0563] Effect of temperature on injectability

[0564] The syringes containing the formulations were stored at 4° C. for 24 hours. After 24 hours, the syringes were removed and the syringeability was immediately measured. The other syringes were stored at 25° C. before measuring the syringeability.

[0565] dispersion

[0566] This test was slightly modified from the test described above in Iteration 1. Each scintillation vial (weighed) was filled with 20 mL of distilled water. 3-4 g of formulation was added to the vial using a syringe. After addition of formulation, the vial was shaken in an orbital shaker at 37° C. and 150 rpm. At predetermined times, the vial was removed and the aqueous layer decanted. The vial was dried at 60° C. for 24 hours. After drying, the weight of the dried vial containing the remaining formulation was recorded.

[0567] Stability studies

[0568] Storage stability

[0569] The formulations were prepared and allowed to stand at room temperature for 24 hours before being filled into HDB syringes (white plunger and needle). The syringes were stored in an incubator at 25°C with the tip pointing upward for varying periods of time. To test the syringeability at different time points, the syringes were removed from the incubator and mounted on a texture analyzer, and the force required to expel the formulation was measured.

[0570] Stability under temperature cycling

[0571] 3-4 g of the formulation was filled into a HDB syringe, stored with the tip facing upward, and then exposed to 4° C. for 24 hours and then to 25° C. for 24 hours. This temperature cycle was repeated for 10 days, and the formulation was then tested for injectability.

[0572] result

[0573] The preparation method affects physical properties, particularly injectability, and therefore the method was standardized as described above. Various preliminary studies on the order of addition of lauric acid and the temperature of lauric acid addition emphasized the need for standardization. (See Example 3.)

[0574] Table 1K. Effect of preparation method on physical properties.

[0575]

[0576] Rheological properties

[0577] Calcium carbonate and calcium phosphate were chosen as fillers / solid particles because of their higher density (higher than sodium chloride and magnesium carbonate). TM In contrast, these formulations prepared by the doctor blade method containing 2.3% aluminum stearate had low viscosity and / or poor rebound behavior. Increasing the aluminum stearate concentration and adding lauric acid improved the formulations. Figure 3 、 Figure 4 、 Figure 5 and Figure 6 .

[0578] Rheological properties of combination filler formulations

[0579] Combinations of fillers were investigated in order to increase density while achieving a similar effect to Teatseal TM Combine calcium carbonate or calcium phosphate (higher density) with magnesium carbonate or sodium chloride. TM In comparison, the calcium carbonate formulation containing 5% magnesium carbonate and the calcium phosphate formulation containing 10% NaCl showed good rheological properties. The 55% and 60% sodium chloride formulations also showed good rheological properties and were selected as water-soluble formulations, while the calcium carbonate formulation was an example of a water-insoluble formulation. TM and another commercially available bismuth subnitrate product, Biobloc TM See also Figure 7 and Figure 8 .

[0580] Suspended breast preparation and recovery rate

[0581] The following formulations were prepared by the doctor blade method.

[0582] Table 1L. Formulations used in the suspended breast study.

[0583] Element F6 F7 F8 F9 F10 <![CDATA[MgCO3]]> 5％ 30％ NaCl 10％ 55％ 60％ calcium phosphate 50％ calcium carbonate 50％ Aerosil-200 0.7％ 0.7％ 0.7％ 0.7％ 0.7％ Aluminum stearate 4.3％ 4.3％ 4.3％ 4.3％ Lauric acid 5％ 5％ 5％ Methylene blue 0.3％ 0.3％ 0.3％ 0.3％ 0.3％

[0584] Table 1M. Calculated formulation density.

[0585] preparation <![CDATA[Calculated density (g / cm 3 )]]> <![CDATA[Teatseal TM ]]> 1.90 F6 1.41 F7 1.54 F8 1.32 F9 1.38 F10 1.09

[0586] Recovery of formulations from the hanging breast study

[0587] A hanging breast study was conducted and details of the study can be found in Example 7. In total, nine breasts were used and four prototype formulations were tested. TM and Biobloc TM Use as a standard / control formulation. After infusion of the formulation, collect the fluid dripping from the nipple for gravimetric analysis. One hour after infusion, peel the nipple, then cut and scrape it to recover any residual formulation. When the nipple is removed, any formulation that enters the udder during infusion is also recovered. Flush the formulation with udder secretions or milk and mark it as flushed.

[0588] dripping

[0589] All prototype formulations had good viscosity and dripping occurred in only one udder (F8 30% dripping).

[0590] Squeeze clean

[0591] The formulation was recovered by vigorous squeezing. Most formulations were easily squeezed. The highest amount of prototype formulation recovered by squeezing was nearly 100% in udder-1, while the lowest recovery was 10% in udder-8. Formulation not recovered by squeezing was either in the teat pool or entered the udder during formulation infusion.

[0592] Scratching

[0593] Scraping was performed to recover any remaining formulation that was not expressed from the teat. The highest amount of formulation was recovered by scraping in udder-2, while the lowest amount was recovered by scraping in udder-7. The inability to fully recover formulation from the teat by expression may have contributed to the variability and poor recovery rates in the retention studies described above.

[0594] Break out

[0595] Different amounts of Teatseal TM The formulation flushed out of the udder, indicating that it was injected into the udder, although the teat was clamped at the base during injection. This also has important implications for retention studies and the efficacy of teat sealant formulations. Figure 9 .

[0596] Storage stability studies

[0597] The storage stability of the formulations tested in the hanging breast study of Example 7 showed physical instability when stored at 25°C. The injection force of the calcium carbonate and magnesium carbonate formulations increased to greater than 4500g at 25°C. Therefore, these two formulations were not used in subsequent studies. The other formulations did not harden upon storage. See Example 1 Figure 10 and Figure 11 .

[0598] Effect of temperature cycling (4 and 25°C) on storage stability

[0599] Temperature cycling did not affect the injectability of the calcium phosphate and sodium chloride (55%) formulations, but the 60% sodium chloride formulation showed slightly improved injectability compared to day 0. Figure 12 .

[0600] Iteration 3 - Laboratory Trials of the Suspended Breast Study

[0601] Based on the results of Iteration 2, it was decided to move forward with the sodium chloride and calcium phosphate formulations. These formulations were laboratory tested and sent for antimicrobial testing. The description and results of this microbial testing are shown in Example 5.

[0602] Sodium chloride and calcium phosphate preparations

[0603] Several sodium chloride and calcium phosphate formulations were tested for their rheological properties and storage stability.

[0604] Table 1N. Syringability and rheological properties.

[0605]

[0606] All formulations contained 4.3% aluminum stearate and 0.7% Aerosil. ND = Not Determined.

[0607] Storage stability at 25°C

[0608] The syringes containing the formulations were stored with the tip pointing upwards, assuming that if phase separation occurred, it would be visible when measuring syringeability. The syringes were stored at 25°C and the formulations were observed for phase separation at predetermined time points and tested for syringeability. No phase separation (formation of an oily layer) occurred. All prototype formulations did not show any signs of hardening during storage for up to six weeks. Figure 13 .

[0609] Effect of temperature on injectability

[0610] The prototype and commercial formulations were stored at 4°C for 24 hours and tested for syringeability immediately after removal from the refrigerator. All controlled formulations showed good syringeability at room temperature. However, the prototype formulation had higher syringeability due to the hardening of the formulation at a lower temperature. Due to the presence of air bubbles in the syringe, the Teatseal TM showed unreliable results, while Biobloc TM The formulation showed hardening during storage at 4°C. TM and Biobloc TM The formulations were filled into HDB syringes and tested for injectability after storage at 4°C for 24 hours. TM and Biobloc TM The preparation also showed hardening of the preparation at 4°C.

[0611] Table 1O. Injectability of commercial and prototype formulations after storage at 4°C for 24 hours.

[0612] preparation Temperature (℃) syringe Injection force (g) Teatseal RT Teatseal 650 Teatseal 4 Teatseal 1125 Teatseal RT HDB 3500 Teatseal 4 HDB 5500 Teatseal RT HDB 3700 Teatseal 4 HDB 6200 BioBloc RT BioBloc 2850 BioBloc 4 BioBloc 5500 BioBloc RT HDB 3900 BioBloc 4 HDB 5600 55% NaCl + 2.5% LA-RT RT HDB 3400 55% NaCl + 2.5% LA-4C 4 HDB 8900 50% CP + 5% LA-RT RT HDB 3300 50% CP + 5% LA-RT 4 HDB 105000 35% CP + 25% NaCl + 5% LA-RT RT HDB 3100 35% CP + 25% NaCl + 5% LA-RT 4 HDB 7500

[0613] All prototype formulations contained 4.3% aluminum stearate and 0.7% Aerosil. RT = Room Temperature

[0614] Dispersion test

[0615] Calcium phosphate and sodium chloride formulations were tested for up to six weeks. After each time point, gravimetric analysis was performed to calculate the loss of water-soluble salts. Calcium phosphate formulations containing 25% NaCl lost approximately 100% of their water-soluble salts after six weeks, while formulations containing 55% NaCl showed the greatest loss of approximately 30-40%, and formulations containing 60% NaCl showed a 50% loss after six weeks. These results point to a formulation that is a slow-eroding sealant formulation.

[0616] Table 1P. Loss of water-soluble components (NaCl) of calcium phosphate formulations in the dispersion test (%).

[0617]

[0618]

[0619] All formulations contained 4.3% aluminum stearate and 0.7% Aerosil.

[0620] Table 1Q. Loss of NaCl formulations in the dispersion test (%).

[0621]

[0622]

[0623] All formulations contained 4.3% aluminum stearate and 0.7% Aerosil.

[0624] Comparison of NaCl and KCl formulations (rheological properties)

[0625] Some preliminary trials were conducted using a ground potassium chloride (KCl) formulation as an alternative soluble salt. The KCl formulation exhibited similar rheological properties compared to the NaCl (55%) formulation, suggesting it is a suitable replacement for NaCl. Figure 14 and Figure 15 .

[0626] Hanging Breast Study

[0627] Based on laboratory and microbiological testing, the following formulations were selected for use in the hanging breast study described in Example 7.

[0628] Table 1R. Formulations selected for the breast suspension study.

[0629] Element F1 F2 F3 F4 F5 NaCl 55％ 10％ 25％ 10％ calcium phosphate 50％ 35％ 50％ 60％ Aerosil-200 0.7％ 0.7％ 0.7％ 0.7％ 0.7％ Aluminum stearate 4.3％ 4.3％ 4.3％ 4.3％ Lauric acid 2.5％ 5％ 5％ 5％ 5％

[0630] Summarize

[0631] Based on laboratory-based trials, udder hang trials, and some preliminary data from retention studies, the use of high concentrations of sodium chloride, alone or in combination with calcium phosphate suspended in oil gelled with aluminum stearate and fumed silica (Aerosil-200), and containing lauric acid, shows promise as a teat sealant formulation that overcomes the current reliance on bismuth subnitrate. Potassium chloride may be a suitable alternative to sodium chloride. The idea of ​​using soluble fillers / solid particles is indeed novel and opens up the possibility of designing slow-eroding teat sealants, thereby overcoming the need for clearing out at the end of the dry period.

[0632] The rheological properties (shear thinning, rebound) of such formulations can be controlled by varying the percentage of sodium chloride and / or calcium phosphate and the concentration of aluminum stearate and Aerosil-200. The formulations can be adjusted to alter the rate of dispersion in water, suggesting that it may be possible to design formulations to erode / disperse at a tailored rate within the nipple.

[0633] Short-term stability studies demonstrated that, unlike magnesium carbonate and calcium carbonate based formulations, sodium chloride and calcium phosphate formulations were physically stable (as assessed by syringeability and rheology).

[0634] These teat sealant formulations that do not contain bismuth subnitrate have antimicrobial activity against mastitis-causing bacteria in vitro (see Example 5).

[0635] Example 2 - Teatseal TM Assessment

[0636] The commercially available Teatseal was evaluated TM by:

[0637] -Determine batch-to-batch variations in rheological properties.

[0638] - Understand the effect of excipients on rheological behavior.

[0639] - Provide some guidance on the preferred rheological properties of internal teat sealants.

[0640] Rheometry

[0641] An internal teat sealant formulation was prepared by the beaker method according to PCT Publication No. WO 2010 / 065747 A2 (the entire contents of which are incorporated herein by reference). Commercially available Teatseal was tested in various ways. TM , Rheology of the internal treatment sealant formulations and different combinations (Table 2A):

[0642] - Shear thinning behavior, i.e. the relationship between viscosity and shear rate.

[0643] - Injection simulation, which simulates injection with a high initial shear rate, followed by a continuous application of low shear rates to understand the recovery rate after injection.

[0644] The rheological properties of the formulations were tested using a Discovery Hybrid Rheometer HR-3 (TA instruments). Samples (4-5 g) were placed on the rheometer plate. The cone / plate geometry was: 2°, 60 mm, and a gap of 500 μm. The temperature was fixed at 32°C.

[0645] Table 2A. Formulations containing bismuth subnitrate.

[0646] Serial number Fillers (65%) Aerosil-200(%) Aluminum stearate (%) 1 bismuth subnitrate --- --- 2 bismuth subnitrate 1 --- 3 bismuth subnitrate --- 2.3 4 --- 1 2.3

[0647] All formulations contained paraffin oil.

[0648] Injectability

[0649] Batches and in-house formulations were also evaluated using a texture analyzer to measure the force and work required to expel the formulation from a syringe. The syringe plunger advances a distance of 25 mm at a constant speed of 2 mm / s to expel the formulation in 12.5 seconds. The analyzer continuously measures force, providing a graph of force versus time.

[0650] Particle size and shape analysis

[0651] Since the rheology of concentrated suspensions is affected by the size, shape and surface characteristics of the suspended particles, size and shape were assessed by scanning electron microscopy (SEM).

[0652] Teatseal TM (1 g) was added to a Falcon tube (15 mL), followed by the addition of hexane (10 mL) to remove the paraffin oil. TM Mix with hexane and shake (by hand) and centrifuge at 3000rpm at 25 ℃ for 30 minutes. After centrifugation, remove the supernatant and repeat this step three times. The extracted bismuth subnitrate is dried overnight at room temperature. Scanning electron microscopy (SEM) is used to analyze the particle size and shape of bismuth subnitrate. Commercially available bismuth subnitrate (Sigma-Aldrich) is used as a reference.

[0653] result

[0654] Teatseal TM Batch rheology

[0655] The six batches showed considerable batch-to-batch variability in their shear-thinning behavior ( Figure 16In injection simulations, there were also differences in the behavior of the batches, particularly in the "rebound" at reduced shear rates, i.e., gelation behavior ( Figure 17 ).

[0656] Injectability

[0657] The maximum force between different batches was from 1500 to 3300 g weight ( Figure 18 ).

[0658] Particle size and shape analysis

[0659] Bismuth subnitrate is commercially available and is from Teatseal TM The extracted bismuth subnitrate showed a range of particle sizes (10-40 μm) and rod-shaped particles.

[0660] Teatseal TM Effects of various excipients in

[0661] Teatseal TM Research was conducted to understand the impact of each ingredient on Teatseal TM Different combinations were made and the rheological properties were tested (Table 2B).

[0662] Table 2B. Rheological properties of formulations containing bismuth subnitrate.

[0663]

[0664] 5 = Internal teat sealant formulation.

[0665] Flow behavior - shear thinning behavior

[0666] Flow behavior evaluates the change in viscosity with increasing shear rate. When undisturbed, the formulation should be viscous, but when shear is applied, the formulation should thin out and flow like a liquid so that it can be easily discharged from a syringe. In other words, when shear is applied, the sample exhibits shear thinning. Aerosil-200 and aluminum stearate without bismuth subnitrate exhibited poor initial viscosity (Table A2). Bismuth subnitrate alone in paraffin oil showed a higher, but still poor, initial viscosity. Bismuth subnitrate added to oil using an aerosol showed an increased viscosity (84 Pa.s). The internal teat sealant formulation showed good initial viscosity (Table A2), comparable to the commercially available Teatseal TM Equivalent: 180Pa.s, while commercially available Teatseal TM (143Pa.s). This difference belongs to Teatseal TM The batch-to-batch variability is large and may be caused by multiple factors: preparation method, storage time, and particle properties of bismuth subnitrate.

[0667] Injection simulation

[0668] This test is to simulate the injection of a formulation into the nipple. In this test, a high shear rate (550 (1 / s)) is initially applied for a few seconds, and then a low shear rate (10 (1 / s)) is applied a few seconds later to simulate the situation in the nipple shortly after administration. At low shear rates, the formulation thickens quickly (so-called "rebound") so that it remains in the nipple. Preferably, rebound should not occur immediately, but time should be allowed for the formulation to diffuse into the gaps in the nipple pool. If the formulation does not rebound, it will drip from the nipple. Samples without bismuth subnitrate and aluminum stearate showed poor rebound (Table A2). This test shows that the combination of aluminum stearate and bismuth subnitrate is important for rebound performance.

[0669] Example 3 - Preparation of an Experimental Teat Sealant Formulation Containing Lauric Acid

[0670] In this example, the inventors describe the general preparation of a teat sealant composition / formulation that does not contain bismuth salts, but otherwise has the same functional characteristics as Teatseal. TM similar.

[0671] The present inventors have discovered that a teat sealant formulation that does not contain bismuth metal can be formulated by the following steps:

[0672] - A water-insoluble gel is formed by mixing an oily carrier (such as paraffin oil) with a thickener (gelling agent) (such as aluminum stearate).

[0673] - Adding another thickener to the gel, such as fumed silica (Aerosil-200), to change the flow behavior of the gel.

[0674] - Solid particles (fillers) are added to the gel to obtain a paste containing coarse solid particles / fillers. The particles can be water-soluble or water-insoluble. Various solid particle types can be used, including sodium chloride (which is water-soluble), potassium chloride (which is water-soluble), sucrose (which is very soluble in water), mannitol (which is soluble in water), dicalcium phosphate dihydrate (calcium hydrogen phosphate dihydrate) (which is almost insoluble in water), magnesium carbonate (which is slightly soluble in water), calcium carbonate (which is almost insoluble in water), etc. The solid particles can be ground to a specific average particle size. The particles change the rheological properties of the formulation and can also provide antimicrobial performance.

[0675] - Addition of an additional thickener, such as lauric acid (which is practically insoluble in water), which provides antibacterial properties to the paste.

[0676] By varying the percentage of sodium chloride and / or calcium phosphate, as well as the concentrations of aluminum stearate and fumed silica (Aerosil-200), the rheological properties (shear thinning, rebound) of such formulations can be controlled. The formulations can be adjusted to alter the rate of dispersion in water, suggesting that it may be possible to design formulations to erode / disperse at a tailored rate within the nipple.

[0677] Teat sealant formulations are generally prepared as follows.

[0678] Preheat the oven to 120°C. Add paraffin oil (Sigma-Aldrich) and aluminum stearate (Strem Chemicals, Inc.) to a beaker and mix. Heat the mixture at 120°C for 40 minutes to melt the aluminum stearate. Remove the beaker from the oven and mix with a spatula to form a gel. Cool the gel to room temperature within 30-40 minutes. Weigh Aerosil-200 (Chemiplas (NZ) Ltd) and place it on a glass plate. Use a spatula to mix the gel and Aerosil-200 on a glass plate. Add solid particles / fillers and mix thoroughly with a spatula to obtain a smooth suspension / paste containing coarse fillers / solid particles.

[0679] The suspension was added to a beaker and melted lauric acid (45°C) was added and stirred. The suspension was returned to the plate and stirred with a spatula for a few minutes to break up any lauric acid lumps. The formulation was allowed to stand in the beaker at room temperature for 24 hours and then tested for its rheological properties.

[0680] Various coarse fillers / solid particles can be used: sodium chloride, potassium chloride, dicalcium phosphate dihydrate etc. Use calcium hydrogen phosphate dihydrate (BP level, Source-Sigma-Aldrich). NaCl (pharmaceutical grade, Source-Dominion Salt Limited, NZ) is ground for 1h (speed-600rpm) and sieved (90 μm) in a planetary ball mill (Retsch PM-100, ball size 9mm). The size of the NaCl particles ground by electron microscopy, and found <10 μm (average size).

[0681] It was found that the method and timing of lauric acid addition affected the rheological properties of the formulation. Therefore, various methods of adding lauric acid were investigated, as described below.

[0682] Method 1 - Add lauric acid to oil containing aluminum stearate and heat at 120°C. After melting the lauric acid and aluminum stearate, cool the mixture to room temperature. Mix the mixture with the filler on a glass plate and stir with a spatula until a smooth preparation is obtained.

[0683] Method 2 - Lauric acid was melted in an oven at 45°C and added to the suspension (room temperature), and the lauric acid lumps were then broken up with a spatula.

[0684] The formulations were allowed to stand at room temperature for 24 hours and then tested for their rheological properties.

[0685] Figure 19 The flow behavior of two formulations of identical composition is shown.

[0686] Figure 20 The effect of the method of lauric acid addition on the rheological properties during injection simulation tests is shown.

[0687] Adding lauric acid to room temperature formulations vs. warm formulations

[0688] Method 3 - Add the melted lauric acid to the room temperature formulation and stir the formulation with a spatula. (See Methods section).

[0689] Method 4 - To prepare this formulation, follow the above steps (Methods section).Put the formulation in a beaker and add lauric acid (solid) to the formulation and heat the mixture in an oven at 45°C for 30 minutes and then mix with a spatula in the beaker (warm formulation).

[0690] The formulations were allowed to stand at room temperature for 24 hours and then tested for their rheological properties.

[0691] Figure 21 Shown is the effect of adding lauric acid to room temperature formulations vs. warmed formulations on the rheology (viscosity vs. shear rate) of the formulations - blue (room temperature formulation), orange (warmed formulation).

[0692] Figure 22 The effect of adding lauric acid to room temperature formulations vs. warmed formulations on rheological properties during injection simulations is shown.

[0693] Example 4 - Rheological Properties of Teat Sealant Formulations

[0694] In this example, the inventors show that the teat sealant formulations described in the above examples have similar properties to the commercially available bismuth-containing teat sealant product, Teatseal. TM Similar rheological properties.

[0695] Figure 23 Shown with Teastseal TM and Biobloc TM Comparison of the flow behavior of various experimental teat sealant formulations.

[0696] Figure 24 Shown with Teastseal TM and Biobloc TMCompared to injection simulations of various experimental teat sealant formulations.

[0697] Figure 25 Shown with Teastseal TM and Biobloc TM Comparison of the flow behavior of various experimental teat sealant formulations.

[0698] Figure 26 Shown with Teastseal TM and Biobloc TM Compared to injection simulations of various experimental teat sealant formulations.

[0699] Figure 27 Shown with Biobloc TM In comparison, the injectability of teat sealant formulations was tested.

[0700] Figure 28 Shown with Teastseal TM and Biobloc TM Comparison of the flow behavior of various experimental teat sealant formulations containing calcium carbonate.

[0701] BP grade calcium carbonate preparations

[0702] Figure 29 Shown with Teastseal TM and Biobloc TM Compared to injection simulations of various experimental teat sealant formulations containing calcium carbonate.

[0703] Figure 30 Shown with Biobloc TM Comparison of the injectability of various experimental teat sealant formulations containing calcium carbonate.

[0704] Figure 31 Shown with Teastseal TM and Biobloc TM Comparison of the flow behavior of various experimental teat sealant formulations containing dibasic calcium carbonate.

[0705] Figure 32 Shown with Teastseal TM and Biobloc TM Compared to injection simulations of various experimental teat sealant formulations containing dibasic calcium carbonate.

[0706] Figure 33 Shown with Biobloc TM Compared to the injectability of the experimental teat sealant formulation, which was obtained from BioBloc TMDelivered with a syringe or a white HDB syringe.

[0707] Figure 34 Biobloc shown using its own syringe TM Injectability.

[0708] Figure 35 Biobloc shown using a white HDB syringe TM Injectability.

[0709] Figure 36 Shown is the injectability of the experimental teat sealant formulation delivered using a white HDB syringe.

[0710] Figure 37 Shows the use of Biobloc TM Injectability of the experimental teat sealant formulations delivered by syringe.

[0711] Summarize

[0712] Flow plots of viscosity versus shear rate are presented, along with simulated plots where the shear rate starts high (simulating injection), then decreases (simulating sedimentation), and finally reaches zero (simulating solidification in the nipple pool).

[0713] All of these were compared to two commercially available teat sealants (TeatSeal TM and BioBloc TM ) for comparison.

[0714] The behavior of the inventor's lead formulation (50% CaHPO4 + 10% NaCl + 4.3% aluminum stearate + 0.7% fumed silica + LA added last) in water (which simulates milk) and in a syringe (which simulates a nipple) is as follows: Figure 39 and 38 shown.

[0715] in conclusion

[0716] The flow properties (rheology) of the teat sealant formulation were similar to commercially available teat sealants.

[0717] Example 5 - Antimicrobial Activity of Teat Sealant Formulations

[0718] Summarize

[0719] This example shows that commercially available bismuth-containing Teatseal TM The present inventors have developed a novel bismuth subnitrate-free teat sealant formulation / composition that has antimicrobial activity against mastitis-causing bacteria in vitro.TM Antimicrobial properties were demonstrated against Streptococcus uberis (S. uberis), but it had no effect on the growth of Staphylococcus aureus (S. aureus) or Escherichia coli (E. coli). Sodium chloride (NaCl) at 55% w / w with lauric acid (LA) at 5% w / w killed all three bacteria. Potassium chloride (KCl) at 50% w / w with 2.5% w / w LA killed both bacteria tested: S. uberis and E. coli. Calcium phosphate (dicalcium phosphate dihydrate, which is calcium hydrogen phosphate dihydrate) with several concentrations of NaCl and 5% w / w LA killed S. uberis but had no bactericidal effect against E. coli. The in vivo antimicrobial activity of these teat sealant formulations may be as good as or better than the activity observed in vitro.

[0720] background

[0721] Teat sealants are applied to the teats of dairy cows during the dry period to prevent new mastitis infections. Internal teat sealants are typically composed of 65% w / w bismuth subnitrate, aluminum stearate (4.3% w / w), and Aerosil dispersed in a mineral oil vehicle. TM The paste is composed of fumed silica (0.7% w / w) and is in the form of a paste. This antibiotic-free teat sealant paste is applied to the teat canal using a disposable syringe and works by preventing bacteria from entering the udder. It is believed that the paste creates a physical barrier between the udder and the environment, thereby preventing bacteria from entering.

[0722] The objective of this work was to evaluate the in vitro antimicrobial activity of teat sealant formulations against the mastitis-causing bacteria Staphylococcus aureus, Streptococcus uberis, and Escherichia coli. The antimicrobial activity of the pastes was determined by counting viable bacteria in culture over time, either exposed to the paste beforehand (solution kill) or after exposure to the paste suspended in culture (contact kill). Solution kill evaluated whether any agents leached from the paste affected bacterial survival, while contact kill evaluated whether the paste surface affected bacterial survival.

[0723] Materials and methods

[0724] Material

[0725] Bacterial strains Escherichia coli ATCC 10536, Streptococcus uberis ATCC 19436, and Staphylococcus aureus ATCC 6538 were maintained in glycerol stocks (500 μl of culture mixed with 500 μl of 87.5% glycerol) and stored at -80°C.

[0726] method

[0727] Bacterial culture

[0728] S. aureus cultures were grown in TSB (Tryptic Soy Broth), S. uberis cultures were grown in Todd-Hewitt broth (THB), and E. coli cultures were grown in Lysogeny Broth (LB).

[0729] Sterilization of teat sealant preparations

[0730] The syringe, aluminum stearate, Aerosil TM The lauric acid, kaolin, and paraffin oil were sterilized by gamma irradiation. The magnesium carbonate, sodium chloride, and glassware were sterilized by heat sterilization (160°C, 3h).

[0731] Solution kills

[0732] Sterile teat sealant composition (1 g) without bismuth subnitrate was added directly to a separate culture medium (THB or LB) and mixed at 100 rpm at 32°C for a period of 24 or 48 hours. After 24 or 48 hours, a sample of the culture medium was taken for measurement of bacterial counts. The antibacterial activity of the solution was measured by spotting the solution onto a lawn of bacterial cells of Streptococcus uberis or Escherichia coli, or by transferring the solution to a new universal culture medium and inoculating 1 x 10 6 CFU / ml of Escherichia coli or Streptococcus uberis. Bacterial culture medium samples were collected on days 0, 1, and 3 after inoculation for calculation of cell viability.

[0733] Kill on contact

[0734] Sterile teat sealant composition (1.0 g) without bismuth subnitrate was added to a 24-well plate under aseptic conditions, and then 1 ml of culture medium inoculated with bacteria was added to each well and incubated at 32°C and mixed at 100 rpm. For each time point, a sample (50 μl) was taken from each well and serially diluted to 10 with PBS in a 96-well plate. 7 cells / ml (time zero). Then 20 μl of dilution (10 to 10 7 Cells / ml) were plated onto TSA plates and incubated overnight at 37°C to calculate the CFU / ml of each sample. Immediately after inoculation, samples (10 μl) were taken from each universal culture medium and serially diluted to 10 in PBS in a 96-well plate. 7 , to determine the activity at T = 0h. Then 20 μl of the dilution 10 -1 to 10-7 The cells were plated onto TSA plates and incubated overnight at 37°C to calculate the CFU / ml of each sample.

[0735] result

[0736] Solution kills

[0737] Solution killing of E. coli and S. uberis by various paste formulations was evaluated and presented as bacterial cell survival graphs ( Figure 40A -C).

[0738] The antibiotic gentamicin (100 μg / ml) in the culture medium was used as a positive control and was shown to be bactericidal against Escherichia coli and Streptococcus uberis. TM The 24-hour culture medium had no antibacterial effect on the growth of E. coli ( Figure 40A In the culture medium collected 48 hours after exposure to the paste, only 50% NaCl reduced the growth of E. coli compared to the untreated control, as observed on the first day. However, the bacterial counts recovered to the same cell density as the untreated control by day 3 ( Figure 40B ).

[0739] S.uberis was evaluated only in the media exposed to the paste for 48 hours. Media exposed to the formulation containing 50% NaCl and 5% LA for 48 hours killed all S.uberis at the 24 hour time point, and no regrowth of S.uberis was observed at 3 days ( Figure 40C This indicates that for this formulation, the compound has leached from the paste into a solution that is bactericidal against S.uberis. In contrast, for E. coli, the paste containing 50% NaCl was bacteriostatic after 48 hours of exposure to the medium ( Figure 40B ). The 25% MgCO3 + 5% LA formulation had an inhibitory effect on S.uberis for 24 hours, but the cells recovered to the same level as the untreated control and paraffin control before the end of the experiment ( Figure 40C ).

[0740] Kill on contact

[0741] Streptococcus uberis

[0742] Culture medium containing S.uberis was plated onto the paste and bacterial counts were assessed on day 2 and day 5. Figure 41 (AH) shows the bacterial survival rate in the teat sealant formulations over time. Paraffin oil (the vehicle used to make the paste) had no antimicrobial activity ( Figure 41A When lauric acid (LA) was added to paraffin oil, strong antibacterial effects were observed by day 2 when LA was added at 2.5% and 5% w / w, whereas it took 5 days to reach zero bacterial count when LA was added at 1% w / w ( Figure 41A Compared with the control, NaCl alone at concentrations of 20%, 30%, 40% and 55% (w / w) in paraffin oil showed some inhibition of bacterial cell counts on day 2, but on day 5, the bacterial numbers were similar to those of the control ( Figure 41B For all NaCl pastes containing 1%, 2.5% or 5% LA, bacterial cell counts were greatly reduced on day 2 and remained bacteria-free on day 5 ( Figure 41B -F).

[0743] The antimicrobial activity of pastes prepared using dicalcium phosphate dihydrate (CaP) as a filler was also investigated ( Figure 41F These pastes all contained 5% w / w LA and replaced CaP with varying percentages of NaCl; the formulation weight percentages (CaP:NaCl:LA) were 60:0:5, 50:10:5, and 35:25:5. None of the CaP-based formulations showed antimicrobial activity against S.uberis within 5 days of paste exposure. In the same experiment, 55% w / w NaCl with 2.5% w / w LA showed bactericidal activity against S.uberis on day 5 ( Figure 41F ).

[0744] 55% w / w KCl paste alone and in combination with 1%, 2.5% and 5% LA effectively killed all S.uberis on day 1 ( Figure 41G ).

[0745] Kaolin (heavy clay) is used in pharmaceuticals, including topical and oral preparations. Kaolin is a layered silicate mineral with the approximate chemical formula of H2Al2Si2O8 (H2O) and is available in a fine powder form. Kaolin-based formulations without LA and with 1%, 2.5%, or 5% w / w LA were evaluated for their antimicrobial activity against S.uberis. Kaolin-based pastes without LA showed similar bacterial counts to the control, whereas all formulations containing LA showed antimicrobial activity against S.uberis ( Figure 41H ).

[0746] Escherichia coli

[0747] Figure 42 shows the antimicrobial activity of different teat sealant formulations, presented as bacterial counts over time. Gentamycin (an antibiotic known to have antibacterial activity against E. coli) was demonstrated to kill all bacteria on day 0 after mixing ( Figure 42A Teat sealant paste formulations containing 55% or 60% w / w NaCl were able to kill all E. coli on day 15 ( Figure 42A ), while 55% w / w NaCl paste containing 5% w / w LA was able to kill all E. coli at the first time point on day 5 ( Figure 42A Teat sealing pastes containing 50% w / w CaP and 10% w / w NaCl (without or with 5% w / w LA) showed no inhibitory activity against E. coli cell counts over a 20-day exposure period ( Figure 42A ).

[0748] Paraffin oil (PO) without fillers / solid particles but containing 1%, 2.5% or 5% w / w LA showed no antimicrobial activity against E. coli over a 20-day exposure period ( Figure 42B The teat sealant paste containing 55% w / w potassium chloride (KCl) filler showed no antimicrobial activity against E. coli, however, after the addition of 2.5% w / w LA, the bacterial cell count dropped sharply to zero counts on day 5 ( Figure 42B ).

[0749] Staphylococcus aureus

[0750] Staphylococcus aureus is a Gram-positive bacterium that was completely inhibited by gentamicin on day 0, as shown in Figure 43. Paraffin oil had no effect on the growth of Staphylococcus aureus ( Figure 43A ). Commercially available teat sealant Teatseal TM and prototype teat sealant formulation pastes (containing only 50%, 55%, or 60% w / w NaCl or 25% w / w MgCO3 with 5% w / w LA) had no inhibitory activity against S. aureus ( Figure 43A and B). The prototype teat sealant formulation of 50% w / w NaCl with 5% w / w LA showed strong antimicrobial activity with zero colony counts on day 5 ( Figure 43A The paste formulations containing 50% w / w CaP and 10% w / w NaCl (without LA and with 5% w / w LA) had no inhibitory activity against S. aureus ( Figure 43B ).

[0751] Summary of results

[0752] Table 5A. Summary of solution kill in culture media after 48 hours of exposure to different teat sealant prototype paste formulations.

[0753] Nipple sealant Streptococcus uberis Escherichia coli Gentamycin + + paraffin oil - - <![CDATA[MgCO3 25% / LA 5%]]> - - NaCl 50% - - NaCl 50% / LA 5% + - <![CDATA[TeatSeal TM ]]> - -

[0754] (-)<3log decrease (+)>3log decrease

[0755] Table 5B. Summary of contact killing of bacteria by different teat sealant prototype paste formulations.

[0756]

[0757]

[0758]

[0759] (-)<3log decrease (+)>3log decrease

[0760] in conclusion

[0761] This example shows that commercially available Teatseal TM The new bismuth subnitrate-free teat sealant prototype developed by the present inventors has antimicrobial activity against mastitis-causing bacteria in vitro.

[0762] A formulation containing 55% w / w sodium chloride (NaCl) and 5% w / w lauric acid (LA) killed all three bacteria.

[0763] Formulations containing calcium phosphate, several concentrations of NaCl, and 5% w / w LA killed S.uberis but not E. coli.

[0764] Based on laboratory work and on clinical trial results, two lead formulations were selected for noninferiority clinical field studies (i.e., whether the new experimental treatment is unacceptably less effective than the active control treatment).

[0765] Considered performance:

[0766] - Rheology and injectability

[0767] - Antimicrobial activity

[0768] - Residues in milk (based on selected compounds rather than actual residue data)

[0769] -Retention in hanging breasts and live animals

[0770] - Tolerance in living animals

[0771] The following formulations were tested in the non-inferiority field efficacy study.

[0772] These were prepared as described in previous examples.

[0773] Example 6 - Retention of a Novel Teat Sealant Formulation Administered During the Dry Period in Dairy Cows

[0774] summary

[0775] The purpose of this retention study was to determine the quality and proportion of six teat sealant formulations, F1-F6, recovered from teats of four groups of cows scheduled for dry period and compared them with the pioneer bismuth-containing product, TeatSeal TM The study was conducted in autumn on dairy cows in Waikato, New Zealand.

[0776] Six teat sealant test formulations and one commercial product (Teatseal) were administered via intramammary infusion via one tube per quadrant. TM ; Zoetis (TS)) single application. Animals were fed according to normal farm practices to reach the dry period (i.e., restricted intake during the first week of the dry period to encourage milk production to stop). Sixteen cows were selected from approximately 20 cows for dry-off. Cows underwent udder health examinations (rapid mastitis test to check for mastitis, udder palpation to check for inflammation, and poor udder health and teat end scoring to ensure teat end health), and the first 16 recruited had negative RMT, and udder palpation and teat end scores ≤1. Cows were randomly assigned to treatment groups 1 (F1-F3 + TS; 1 hour), 2 (F1-3 + TS; 48 hours), 3 (F1-F3 + TS; 672 hours), or 4 (F4-F6 + TS; 672 hours).

[0777] On day 0, the farmer milked the cows and removed them from the main herd for treatment. Cows were treated in the following order: treatment groups 3, 4, 2, and 1. Treatment group 3 was treated and observed for approximately 1 hour, and then the cows in treatment group 4 were infused with the formulation. Approximately 1 hour after treatment, group 1 was brought back into the shed and, after an ultrasound examination of the teats, the teats were stripped one by one. When the base of the teat was blocked, the injected formulation was carefully squeezed out of the teat until no more secretions (milk and / or teat sealant) were recovered. A second squeeze was collected from treatment group 1, in which milk from the glandular cistern was collected. At 48 hours (2 days) and 672 hours (28 days), the same process used in the squeeze group was used for the remaining treatment groups. All samples were weighed and kept frozen until shipped for further inspection. Ultrasound was also used to examine the teats on days 2 and 28.

[0778] The recovery of the formulations was based on the dry weight of the recovered material after deduction of milk solids. Milk solids deduction was performed in two ways, assuming 10% and 13% milk solids in the milk. There was considerable variability in the amount of teat sealant recovered between formulations, animals, and time. No formulation tested was able to compare favorably with Teatseal. TMConsistently comparable. Teatseal recovered at different times TM There is also a great deal of variation in the amount (and proportion) of milk collected. There are several possible explanations for this variation: Cows are brought into the barn shortly after drying off, which means that the lactation response is triggered. This results in more milk in the teat ducts and may cause product to "leak" due to milk pressure on top of the preparation. It is also relatively difficult to block the teat ducts, hold the collection container, and milk the teats. Physical measures (e.g., fitting a suitable clamp at the base of the teat) can be used to help collect the preparation in this way, and this also helps to milk the teats. Ultrasound examination shows little to no amount of teat sealant in each treated milk cistern / teater duct.

[0779] In summary, all products (including Teatseal TM ) had different recoveries, with no significant difference in teat sealant retention.

[0780] Study Objectives

[0781] The purpose of this study was to determine the quality and ratio of six teat sealant formulations recovered from teats of four groups of cows scheduled for dry period and to compare them with the pioneer product TeatSeal TM (Zoetis) Contrast reserved.

[0782] Study Design

[0783] This was a controlled, randomized study designed to determine the retention of various novel teat sealant formulations in the teats of dry cows and compare them with a commercially available teat sealant (Teatseal TM Sixteen (16) healthy, pregnant, lactating cows of varying ages, scheduled for dry-off, were selected from a commercial dairy farm based on general examination, rapid mastitis test, and udder palpation scores.

[0784] Table 6A. Treatment details

[0785]

[0786] Timeline of events

[0787] Table 6B. Timeline of proposed events.

[0788]

[0789] Research animals

[0790] Species: Cattle

[0791] Breed: New Zealand Friesian cattle

[0792] Number: Sixteen (16)

[0793] Gender: Pregnant female

[0794] Age: Mixed

[0795] Treatment History: No mastitis, previous dry-off treated with dry cow therapy and teat sealants

[0796] Table 6CA. Investigational veterinary products (IVPs) / preparations.

[0797]

[0798]

[0799] Table 6CB. Investigational Veterinary Product (IVP) / Actual Formulation

[0800] Element F1 F2 F3 F4 F5 F6 <![CDATA[MgCO3]]> 25％ 30％ 10％ NaCl 50％ 50％ 25％ Kaolin 45％ Aluminum stearate 4.3％ 5.3％ 4.3％ 5.3％ 4.3％ Aerosil 0.7％ 0.7％ 0.7％ 0.7％ 0.7％ 0.7％ Lauric acid 5％ 5％ 5％ 5％ paraffin oil 65％ 39％ 64.3％ 45％ 44％ 60％

[0801] F4 contained Aerosil-R972, and all other formulations contained Aerosil-200.

[0802] Treatment options

[0803] Treatment and timing

[0804] The cows were milked in the morning according to normal farm practice and were drawn from the herd after being released from the milking shed. They were taken to a yard for treatment.

[0805] IVP and control products were administered on a single occasion (Day 0). IVP was administered via the teat. The treatment schedule for each cow is shown in Table 6D.

[0806] Cows in Treatment Group 3 were treated first and monitored for signs of acute adverse events for approximately one hour. Subsequently, cows in Treatment Group 4 were treated, followed by Cows in Treatment Group 2. These cows were returned to the paddock, where they remained for approximately two weeks (before joining the rest of the herd). Finally, cows in Treatment Group 1 were treated and concluded approximately 6.5 hours after milking was complete, after which they were placed in a pen next to the shed. Approximately 40 minutes later, this group of cows was taken back to the milking shed for milking.

[0807] Table 6D. Treatment plan for each cow.

[0808]

[0809] Breast health

[0810] Umbrella health was assessed prior to Day -1. Each quadrant was observed for swelling and heat as the cow was milked in the milking shed. These observations were made with minimal pressure applied to avoid movement of any IVP / CP around the teat / quadrant.

[0811] Ultrasound procedures

[0812] A portable small animal ultrasound (US) machine was used to determine the amount of teat sealant (if any) in the teat reservoir. To bring the teat into contact with the ultrasound probe, a flexible plastic cup was filled with teat dip and the teat was immersed in the cup. Transverse and / or longitudinal images were captured on video footage and downloaded for viewing.

[0813] Sample collection

[0814] Cows from treatment group 1 were brought back to the milking barn for a 1-hour collection. Cows to be treated were sampled first (1:05-1:18 after product administration). All cows were milked 1:06-1:14 hours after product administration.

[0815] Milking of cows in Treatment Group 2 was performed 48:48-49:36 hours after product administration.

[0816] When the teat base was blocked, the injected formulation was carefully milked from the teat until no more secretions (milk and / or teat sealant) were recovered. The milk, secretions, and teat sealant recovered from each teat were collected in separate plastic tubes. Cows in Group 1 were milked a second time to collect milk / product from the mammary cistern.

[0817] Additional "second squeeze" samples were collected on days 2 and 28.

[0818] Sample analysis

[0819] The sample was dried and the dry mass was determined. Since the formulation contained no volatile components, the difference between the wet and dry mass was due to the aqueous secretions / milk in the wet sample. Assume that the aqueous secretions / milk contained 10% w / w solids. The recovered formulation mass was calculated as follows:

[0820] Recovered mass = dry mass - milk solids

[0821] This was done for the first and second squeeze samples. The percentage recovery was calculated from the mass recovered and the mass applied and compared to the Teatseal TM The percentage of recovery in each cow was compared.

[0822] Statistical analysis

[0823] The average value is calculated based on the raw data results. The raw data is presented in the form of tables and charts. The formula used is:

[0824] Recovery rate relative to the administered amount (%) = (dry mass of the recovered preparation / mass of the injected preparation) x 100

[0825] Relative to Teatseal TM Recovery rate (%) = (Recovery rate of preparation % / Teatseal TM Recovery rate (%) x 100

[0826] The recovery after drying was measured by subtracting the estimated milk solids weight from the dry mass.

[0827] in conclusion

[0828] All products (including TeatSeal TM ) recovery rates varied, with no significant difference in teat sealant retention.

[0829] Other findings

[0830] Due to the large variability and TeatSeal TM retention was lower, and we interpret these results as indicating that there were no clear underperformers among the experimental teat sealants.

[0831] result

[0832] Nipple squeeze

[0833] Graphical nipple squeeze results Figures 50-58 shown.

[0834] On day 2 (48 / 49 hours after treatment), four cows were observed dripping (leaking milk). These animals are listed in Table 6E. "Leaking milk" was observed as the presence of milk drops on the end of the teat.

[0835] Table 6E. Cows observed to be "leaking milk" on day 2 (48 / 49 hours after treatment).

[0836]

[0837]

[0838] The mean weight range for each formulation in each treatment group (time) was: 2.6-2.9 g for the test formulation, and 2.6-2.9 g for the Teatsea TM The content of 4.0-4.1 g was observed (Table 6F).

[0839] The average weight of the preparation recovered from the nipples of the formulation and treatment groups was 0.92 g, including Teatseal TM (Table 6G) The mean values ​​of the test formulations varied over time from 0.12 to 2.93 g, and the Teatseal TM 0.07-6.07g. TM The highest amount of product was recovered in the treated quadrants, consistent with the larger amounts applied. Among the formulations, F2 at 2 days and F3 at 1 hour had the highest recoveries, although the variability was greater for F2 (D2).

[0840] Table 6F. Mean weight (g ± SD) of each formulation administered to each treatment group.

[0841]

[0842] Table 6G. Average weight (g) and range of the formulations recovered 1 hour (first squeeze), 2 days, and 28 days after application.

[0843]

[0844]

[0845] The recovery relative to the amount of formulation administered was variable and ranged from 4.32-40.97% for the test formulations and 0.01% for Teatseal. TM The range was 24.3-73.17% (Table 6G). TM The highest percentage of product recovered was F1 at 1 hour and 28 days (Treatment Groups 3 and 4). On day 2, the highest percentage of product recovered was F1 at 37.82%, followed by Teatseal TM , which is 24.30%.

[0846] Teatseal with a single cow TM In contrast, the average mass of preparation recovered in the cows varied, with values ​​ranging from 15.57% (F4) to 844% (F1, day 2). Overall, the recovery rates were higher in F1-F3 than in F4-F6.

[0847] For the second squeeze, the variation was also large, and in all cases, the product "recovered" (>114%) exceeded the product applied (>114% data not shown). The large variation in this data is difficult to interpret, and therefore this data was not used and is not reported / discussed further.

[0848] Table 6H. Average recovery (%) and range of formulations recovered 1 hour (first squeeze), 2 days, and 28 days after application relative to teat sealant recovery percentage.

[0849]

[0850] Table 6HA. Recovery rate (%) of the formulation at different times after administration.

[0851]

[0852]

[0853] Ultrasound

[0854] A portable small animal ultrasound machine was used to determine the amount of teat sealant in the teat. A score of 0 represented no teat sealant or only a small amount of teat sealant in the teat duct / tea cistern ( Figure 49 A score of 1 indicates a small amount of teat sealant in the teat cistern but not above the teat canal. A score of 2 indicates a small amount of teat sealant at the teat opening (i.e., above the inside of the teat canal, at the bottom of the teat cistern). A teat sealant score of 3 is ideal, filling more than 25% of the teat canal and located at the bottom of the teat cistern, covering the teat canal.

[0855] Cows were ultrasound scanned in the milking shed. Teat sealant scores were recorded for each animal for each teat that was milked (e.g., treatment group 2 at 48 hours, treatment groups 3 and 4 at 28 days). Scores for each formulation (1-6) were averaged and compared to each other and to the control product. Data were presented as arithmetic and / or geometric means, and appropriate statistical analyses were performed.

[0856] Table 6I. Teat sealant ratings and descriptions. (See Figure 49 .)

[0857]

[0858] Ultrasonic waves (US) are captured in video clips, and still images of the cow 302 are obtained from those video clips ( Figures 44-47 ). It is worth noting that for products containing Teatseal TM Unfortunately, using ultrasound to measure the amount of teat sealant in each teat is not feasible. However, with more practice, it may be possible to use ultrasound to determine the presence / absence of teat sealant.

[0859] discuss

[0860] Product recovery varied widely between formulations, animals, and time. No formulation tested matched the market leader, Teatseal. TM In fact, the Teatseal recycled at different times TM The amount (and proportion) of

[0861] Approximately 6.5 hours after milking, cows are "dried off," or treated. Cows are typically dried off in groups to shorten the time between the end of milking and the dry-off treatment. Furthermore, it's uncommon for cows to be brought into the barn soon after drying off (e.g., less than an hour or several days later)—common practice is to place them in a paddock with minimal grass (and possibly hay) to reduce their energy expenditure and thus aid in drying off. Low recovery rates and lack of consistency within the formulation may be due to the lactation response being triggered when the cows are brought into the barn at 48 / 49 hours, resulting in milk dripping. Cows have psychological responses to various noises, smells, or events associated with milking. These responses vary from cow to cow and therefore vary with the time between lactation and milking. Some cows produce milk at the sight of the barn, while others may be triggered by the sound of the milking machine. Without careful behavioral observation over a period of time, it's impossible to pinpoint triggering factors for individual cows with any degree of certainty. Consider milking cows in the yard rather than herding them into the barn where their lactation triggers are activated.

[0862] Because the cows were in the shed multiple times, they also may have been leaking milk. When the cows were brought in for their 48 / 49 hour milking, four cows were observed dripping milk (two, one, and one from treatment groups 2, 3, and 4, respectively). Although the farmer indicated that the cows were ready to dry off, they still seemed to have a little milk (especially on the second day). Figure 48 The results show that even 28 days after dry-off, some variation persists. In future studies, and despite its subjectivity, a scoring system to indicate the extent of udder involution would be useful. It would also be worthwhile to record which cows experience dripping and which quadrants are affected. The cows in the top row may be leaking milk because their teat ducts and mammary cistern are filled with milk.

[0863] Typical milk solids are around 8-12%, with greater variation between breeds (e.g. Jersey cows have higher milk fat and protein than Friesian cows (Holmes et al., 2003). Cows differ in the composition of their milk. Calculations were performed assuming 10% milk solids and repeated assuming 13% milk solids. This did not affect the results much and is because the volume of milk in the first milking sample was not too large.

[0864] The variation may also be due to the difficulty in expressing the teat contents while plugging the teat base and holding the sampling bottle. If a device (e.g., a band) could be applied to the teat base before the cow enters (or approaches) the barn, this could help prevent milk from dripping into the teat ducts and facilitate the process of removing the teat duct contents while avoiding collecting milk from the cistern. This requires careful consideration, as the band / device should not be too tight to irritate the cow, nor too loose to prevent milk from the cistern from dripping into the teat ducts.

[0865] Performing ultrasound examinations of teats in active animals has proven relatively difficult. To avoid applying pressure to the teats, flexible cups filled with fluid are used. It may be possible to use an ultrasound probe without a cup, but the amount of pressure needs to be considered: too much pressure could dislodge the teat sealant, while too little could irritate cows, who do not respond well to having their teats "tickled."

[0866] in conclusion

[0867] Including Teatsea TM Recovery rates varied across all products, with no clear performance in terms of retention. Several changes are recommended for any future work in this area to help potentially reduce some of the variation, such as:

[0868] Express milk immediately before applying the product

[0869] Use a physical device to block the base of the teat and apply it outside the milking shed

[0870] Stay away from the milking shed and milk the teats clean

[0871] Ultrasound examination requires more practice, and in the future it may be used to assess the presence / absence of nipple sealants.

[0872] References

[0873] Holmes,CW,Brookes,IM,Garrick,DM,MacKenzie,DDS,Parkinson,TJ and Wilson,GFMilk production from pasture(Principles and Practices).MasseyUniversity.D.Swain,edited.ISBN 0473 08308 6

[0874] Example 7 - Acute Retention and Recovery of Teat Sealant Formulations in an Ex Vivo Breast Model (Hanging Breast Study)

[0875] Purpose

[0876] The objective of this study was to determine the retention and recovery of a teat sealant formulation approximately 60 minutes following intramammary infusion into the excised udder of a cow.

[0877] Materials and methods

[0878] The study was conducted on two experimental days, separated by a week. On each day, the udders of freshly slaughtered cows were retrieved from a nearby slaughterhouse and transported to the necropsy room. This is a temperature-controlled room set at 18±1°C. Nine udders (18 udders total) were selected each day based on having four functional quadrants and no obvious signs of mastitis. The udder was pierced with a baling needle, and a synthetic rope was passed through the tissue and tied to the frame so that the udder was suspended from the frame in a near-natural position. The length of each teat was then measured to the nearest millimeter, from the base of the teat (at the point of insertion into the udder) to the teat opening.

[0879] After discarding the first three stages of milk, approximately 10 mL of milk was collected from each teat into a single plastic container, and attempts were made to express the remaining milk from each mammary gland pool. The treatments (21OTTS S F1, 21OTTS S F2, 21OTTS SF3, 21OTTS S F4, 21OTTS S F5, and Teatsea) were then randomly assigned to the follicles. TM ) was infused into the nipple (Table 7A; https: / / www.random.org / lists / ). The infusion technique involved holding the base of the nipple with the index finger and thumb of the non-dominant hand to close the nipple cistern and prevent infusion of the therapeutic agent into the mammary cistern. After partially inserting the syringe nozzle (approximately 3 mm) into the streak tubing, the treatment was administered with the dominant hand. The syringe was weighed before and after infusion into the quadrant to determine the mass of the infused product. Table 7B depicts the composition of the trial treatments.

[0880] A separate plastic container was placed under each nipple to collect any secretions / formulation that dripped spontaneously during the first hour or so after infusion. Approximately 60 minutes after infusion, the therapeutic agent was expressed from the nipple as follows: The base of the nipple was gripped with the thumb and index finger of the non-dominant hand, and then repeatedly expressed with the dominant hand until no further product was recovered; the recovered product was collected in a separate plastic container.

[0881] After each papilla was milked, the base of each papilla was closed with Kelly forceps. The papilla was then incised transversely and longitudinally from the dorsal aspect using Kelly forceps. Each papilla cistern was visually inspected and photographed. Representative images of longitudinal cross-sections of each papilla were captured and archived for this study but were not analyzed. Immediately after obtaining photographs of the open papilla, each papilla was scraped, and any recovered contents were placed in individual plastic containers.

[0882] The milk and any preparations flushed from the mammary gland and mammary gland pool during teat incision were collected in a two-liter plastic container. Any visible preparations were placed in a separate plastic container and the milk was discarded.

[0883] All containers were properly identified and weighed before and after sample collection. All samples were stored at -20°C until shipped for analysis.

[0884] Table 7A. Treatment assignments for the 18 breasts.

[0885]

[0886] *FL = Left Front, FR = Right Front, RL = Left Rear, RR = Right Rear.

[0887] + F1 to F5 = novel teat sealant formulations.

[0888] Table 7BA. Composition of the infusion formulation.

[0889]

[0890]

[0891] Calcium phosphate herein refers to calcium hydrogen phosphate dihydrate.

[0892] For balance, paraffin oil was used in all formulations.

[0893] The sample was weighed (ie, wet mass) and dried to constant weight, and the dry mass was determined. Since the formulation contained no volatile components, the difference between the wet mass and the dry mass was due to the aqueous component in the wet sample (Equation 1).

[0894] (1) The mass of water in the sample = wet mass - dry mass

[0895] (Note: dry mass = dry mass of preparation + mass of milk solids)

[0896] Samples collected before treatment infusion were used to determine the percentage of milk solids in the secretions of individual glands (Equation 2). These percentages were used to determine the mass of milk solids in the samples from the corresponding quadrants (Equation 3). The mass of the formulation was then calculated by subtracting the mass of milk solids from the dry mass of each sample (Equation 4).

[0897] (2)

[0898] (3)

[0899] (4) Quality of preparations

[0900] = dry mass - mass of milk solids

[0901] The results are expressed as percentage values ​​relative to the mass of the infused therapeutic agent of the samples collected by dripping, by squeezing, by scraping and by flushing, and the total therapeutic agent considered, according to the following equation:

[0902] (5)

[0903] (6)

[0904] (7)

[0905] (8)

[0906] (9)

[0907] (10)

[0908] Statistical analysis

[0909] Data were analyzed using D'Agostino and Pearson normality tests and presented as mean ± SD and / or median (range) as appropriate. Differences between treatments with respect to teat length, milk solids in milk samples, mass of infused formulation, recovery by dripping, squeezing, scraping, and flushing, and total treatment recovery were analyzed using the Kruskal-Wallis test followed by Dunn's multiple comparison test. Differences between treatments with respect to percent recovery from the teat were analyzed using a GLMM with udder as a random effect and formulation and anterior / posterior teat as fixed effects. Treatments with recovery from the teat greater than or equal to Teatseal were compared using a chi-square test. TM The effect of papilla length on the recovery of the different preparations was determined using linear regression analysis. Values ​​of P < 0.05 were considered significant.

[0910] result

[0911] The nipple lengths of the 18 breasts included in the study ranged from 30 mm to 63 mm, and when the nipples were grouped by treatment, the median lengths did not differ significantly (P = 0.3388; P = 0.001). Figure 59 The estimated solids content of milk samples obtained before infusion ranged from 9.24% to 26.6%, and there was no significant difference in median milk solids content when teats were grouped by treatment (P = 0.1342; P = 0.001). Figures 60-65 ).

[0912] in conclusion

[0913] This study demonstrated that the teat sealant formulation could be recovered approximately 60 minutes after infusion into the teat of an excised cow udder. Most of the infused formulation was recovered by expression followed by scraping and flushing, and the amount of dripping was negligible.

[0914] Further analysis of the results of the previous hanging udder study showed that the recovery rate of posterior teats was lower than that of anterior teats, and that teat length was moderately positively correlated with teat recovery rate (i.e., milking recovery rate + scraping recovery rate; r = 0.4118). Similar to the previous study, the recovery rate of posterior teats was numerically lower than that of anterior teats (94.3% vs. 88.22%), except for those teats treated with 21OTTS S F2. Teat recovery rates were significantly different only in teats treated with 21OTTS S F1 (P = 0.0078). In contrast to the previous hanging udder study, the present study found no correlation between teat length and teat recovery rate (r = 0.0011). Although the inventors had the impression that the teats used in the previous hanging udder study (possibly from heifers) were shorter than the teats used in the current study (possibly from late-lactation cows), teat lengths measured between studies were similar (30 to 55 mm and 30 to 63 mm, respectively). The different results may be due to methodological issues. In a previous study of hanging udders, the length of 27 nipples was measured using photographs of excised, opened nipples and identification labels as a reference. In the present study, 72 nipples were measured using calipers from the inside of the nipple base (at the point of insertion into the udder) to the nipple opening. A method to accurately and consistently measure nipple length needs to be developed to determine the impact of this variable on the recovery of preparations from the nipples.

[0915] In contrast to the previous hanging udder study, in which teat recoveries varied from 1.44% to 126%, more consistent recoveries were obtained with all formulations tested in this study (i.e., 30.96% to 124.57%). This may be attributed to determining the milk solids in each quadrant sample rather than using an untreated quadrant and assuming the milk solids were the same for the other three quadrants of each udder. This may also be related to the significantly longer teats in the udders of the current study (however, see the previous paragraph). Teat recoveries for the formulations tested were also comparable to those obtained with Teatseal. TM The results obtained are similar. Although it was found that when considering at least equal to Teatseal TMWhen the cutoff value for the formulation was met (i.e., ≥95.81%), a significant portion of the anterior papillae had different recoveries for the formulation-infused papillae. However, for example, the three anterior papillae infused with 21OTTS S F1 that did not meet the cutoff mark had recoveries between 94.2% and 94.75%. Similarly, all six anterior papillae treated with 21OTTS S F3 had recoveries between 90.38% and 95.46%. The cutoff benchmark may be viewed as arbitrary, and therefore, interpretations of the differences in papillae recovery with the teatseal formulation should be considered with this limitation in mind. TM Comparison results of recovery rates.

[0916] Some of the infused formulation entered the mammary cistern. However, the recovery rate by flushing in this study (ranging from 0.3% to 51.84%) was less than that in previous studies of hanging udders (ranging from 23.4% to 87.87%). This may be due to the use of partial tip insertion and significantly longer teats in this study, compared with full tip insertion and significantly shorter teats in previous studies. Although syringe tip insertion length appears to have an effect on the recovery rate of infused formulation from the mammary cistern, further studies are needed to test this hypothesis.

[0917] After squeezing out, some preparation may still be present in the teat cistern, e.g. Figure 66 As shown in the photo. Preparation 21OTTS SF5 and Teatseal TM The formulations exhibited a more similar bioadhesion pattern, with significantly more formulation remaining in the teat sump compared to the other tested formulations. This was confirmed by measuring scrape recovery. Different rheological responses to shear stress may have influenced the observed results. The distribution of formulations in the teat sump reported in this study does not reflect what may occur in undisturbed teats or teats subjected to udder movement in live cows.

[0918] In conclusion, the recovery of teat sealant formulations from teats was comparable to that of Teatseal for both anterior and posterior teats. TM The recovery rates of the samples were comparable. The recovery rates obtained by squeezing, scraping and flushing accounted for the majority of the recovery rates; the recovery rates obtained by dripping were negligible.

[0919] Example 8 - Pilot-Scale Local Tolerance Study of Two Novel Teat Sealant Formulations in Dry Cows

[0920] summary

[0921] The scope of this study was to investigate the local and systemic tolerability of two teat sealant formulations (Teat Sealant 1 and Teat Sealant 2) in dairy cows during the dry period. On study day (SD) -1, a total of 10 lactating cows (age: mean = 3.1 years; minimum = 2 years; maximum = 6 years) were enrolled in the study and divided into two study groups of 5 animals each. On SD 0, before dry off, two quadrants of each cow in study group 1 were treated with teat sealant 1 (TS1) and two quadrants of each cow in study group 2 were treated with teat sealant 2 (TS2). All remaining quadrants were treated with veterinary (adus.Vet) OrbeSeal TM (consisting of 65% bismuth subnitrate in a mineral oil vehicle). Local tolerance was assessed once daily at 1 hour, 4 hours, and 12 hours after application during the first week and once daily during the second week. Systemic tolerance was assessed daily by general observation, including rectal temperature measurement. Cows were euthanized on SD 14 following the end-of-study examination. Udder samples were removed and fixed in 10% formalin for histopathological evaluation.

[0922] Two adverse events occurred. Cow I 363 developed severe clinical mastitis in her right hind udder quadrant on SD 2, which required concurrent treatment. TM The quadrant was treated, and a relationship with Staphylococcus aureus detected in the corresponding milk sample after arrival cannot be ruled out. Cow I 366 developed clinical mastitis in the left anterior quadrant on SD 4. Both affected quadrants were sealed with Orbeseal on SD 0. TM Apart from three cows that experienced milk dripping after the dry period, no other findings were observed during the local tolerance evaluation.

[0923] Histopathologically, TS1 and 2 or Orbeseal TM Intramammary administration of did not reveal any local toxic effects related to the investigational veterinary product (IVP). After two weeks, all cows were still clearly lactating. Nevertheless, two cows (I360 and I366) showed chronic active inflammation in one or both udder quadrants, which cannot be excluded as occurring before treatment and / or secondary to an exacerbation during the dry period. Reactive cell proliferation was seen in the associated lymph nodes. Cow I363 developed a subacute abscess in one quadrant, which was consistent with the clinical findings during treatment. Many cows did not show major histopathological findings, but had an increase in granulocytes and / or mononuclear cells or debris in the mammary alveoli, suggestive of a udder degeneration process.

[0924] In conclusion, TS1 and TS2 were well tolerated both systemically and locally.

[0925] Research Scope

[0926] The scope of this study was to investigate the local and systemic tolerability of two teat sealant formulations (TS1 and TS2) in dairy cows during the dry period.

[0927] Animals and methods

[0928] On study day (SD)-1, a total of 10 lactating cows (age: mean = 3.1 years; minimum = 2 years; maximum = 6 years) were enrolled in the study and divided into two study groups of 5 animals each. During the acclimation period and at enrollment approximately 1 month later, milk samples (SD-1) were collected for determination of somatic cell counts. Bacteriological examinations were also performed.

[0929] Female Holstein cows (5 cows / experiment) were treated intramammarily once with TS1 or TS2 at a dose volume of 5 g per quadrant for an observation period of approximately 2 weeks. Cows were randomized so that 2 quadrants received the test substance and the other 2 quadrants received a single dose of 4 g of Orbeseal as a reference substance. TM .

[0930] On SD 0, before dry-off, two quadrants of each cow in Study Group 1 were treated with TS1, and two quadrants of each cow in Study Group 2 were treated with TS2. All remaining quadrants were treated with OrbeSeal®. TM Treatment. Local tolerance was assessed once daily at 1 h, 4 h, and 12 h after application during the first week and once daily during the second week. Systemic tolerance was assessed daily by general observation, including rectal temperature measurement. Approximately 4 h after treatment on SD 0, cows were moved from the tethered barn to a nearby free-range barn.

[0931] Cows were euthanized after the end-of-study examination on SD 14. Udder samples were removed and fixed in 10% formalin for histopathological evaluation.

[0932] result

[0933] At inclusion (on SD-1), quadrant assignment to teat sealant formulation or reference article treatment was determined by lottery according to the study outline. The following treatment patterns were determined according to Table 8A.

[0934] Table 8A. Treatment modalities.

[0935]

[0936] Table 8B. Body weight determination [kg].

[0937] Animal ID Weight [kg] I359 650 I360 582 I361 624 I362 550 I363 683 I364 675 I365 746 I366 506 I367 604 I368 568

[0938] Milk sampling

[0939] Milk samples were collected according to the protocol for determination of somatic cell counts (during adaptation and at SD-1) and for bacteriological examination.

[0940] Table 8C. Bacteriological examination.

[0941] Animal ID Left anterior quadrant Left posterior quadrant right front quadrant Right posterior quadrant 1359 Negative hemolytic Staphylococci Negative Negative 1360 Negative Negative Negative hemolytic Staphylococci 1361 hemolytic Staphylococci Negative Negative Negative I 362 Negative Negative Negative Negative I 363 hemolytic Staphylococci hemolytic Staphylococci hemolytic Staphylococci hemolytic Staphylococci I 364 Negative Negative hemolytic Staphylococci hemolytic Staphylococci I 365 Negative Negative Negative Negative I 366 Staphylococcus aureus Negative Negative Negative I 367 Negative hemolytic Staphylococci Negative Negative I 368 Negative Negative Negative Negative

[0942] The main mastitis pathogen (Staphylococcus aureus) was identified only in the left anterior quadrant of cow I 366. Since S. aureus is sensitive to amoxicillin, it was isolated using Synulox LC Plus TM Treat that quadrant. All other quadrants are free of major pathogens.

[0943] Table 8D. Determination of somatic cell count [x1000 cells / mL].

[0944]

[0945] Even if the inclusion criteria defined otherwise, animals with a somatic cell count in the quadrant greater than 200,000 cells / mL were included in the study. Upon arrival and after inclusion in the study, all quadrants and milk from each quadrant showed no clinical abnormalities. The main mastitis pathogen (Staphylococcus aureus) was detected only in the left anterior quadrant of cow I 366 and was isolated using Synulox LC Plus. TM This quadrant was treated. All other quadrants were free of major pathogens. Based on the clinical picture and the results of bacteriological examinations upon arrival and subsequent treatment, it was decided to include all quadrants in the study despite the increase in somatic cell counts.

[0946] Daily overall health observation

[0947] During the acclimation period and the study period until the dry period, milk production was measured twice daily. General health was checked daily during feeding and cleaning. Severity scores (SV) were assessed at least once daily during the study period. Cow 1363, from SD 3 to SD 8, was assigned to SV3 because she developed severe acute mastitis, which required concurrent treatment. All other cows were assigned to SV2.

[0948] Physical examination

[0949] Physical examinations were performed on SD-1 and before necropsy on SD 14. According to Table 8E, the following findings were observed.

[0950] Table 8E. Physical examination.

[0951]

[0952] Investigational Veterinary Products (IVP) - TS1 and TS2

[0953] Table 8F. Teat Sealant 1 (TS1) Formulation.

[0954]

[0955]

[0956] Table 8G. Teat Sealant 2 (TS2) Formulation.

[0957] Element TS2 <![CDATA[MgCO3]]> NaCl 50％ Kaolin Aluminum stearate 5.3％ Aerosil 0.7％ Lauric acid 5％ Liquid paraffin 39％

[0958] application

[0959] Intramammary application of IVP and reference articles was performed according to the study guidelines. In each case, the full dose was applied without any loss. Teat sealant 1 (TS1) was difficult to apply and required two hands to push the entire contents into the teat. Teat sealant 2 (TS2) was applied in the same manner as OrbeSeal. TM Thereafter, application (TS2) appeared to cause discomfort in the area around the nipple.

[0960] Local and systemic tolerance assessment

[0961] During the local tolerance assessment, all breast quadrants were inspected for signs of inflammation (e.g., redness, swelling, warmth, pain). The following observations were made:

[0962] -I 359: 1 drop of milk in SD

[0963] -I 360: milk in SD 0 (12h) and SD 1

[0964] -I 362: 1 drop of milk in SD

[0965] -I 363: Mastitis in the right posterior quadrant on SDs 2, 3, 4, and 5. Inflammation spreads from the right posterior quadrant to the right anterior quadrant on SDs 10 and 14

[0966] -I 366: Swelling in the left anterior quadrant on SDs 4 and 5. Swelling spread from the left anterior quadrant to the left posterior quadrant on SD 14.

[0967] Rectal temperatures were measured daily. Some cows had elevated rectal temperatures after the dry period, but these were not clinically significant, with the exception of cow I 363, which developed a transient fever due to severe clinical mastitis.

[0968] Adverse events

[0969] Two adverse events occurred during the study period.

[0970] SD 2, I 363 (moderate adverse event): severe swelling and pain in the right posterior breast quadrant.

[0971] SD 4, I 366 (mild adverse event): swelling of the left anterior breast quadrant, no pain, overall well-being was not disturbed.

[0972] Clinical Summary and Concomitant Therapy

[0973] Cow I 363 developed severe mastitis and had to be treated.

[0974] SD 2: severe swelling and pain in the right posterior quadrant, rectal temperature: 39.7°C, treated with Septoman TM Topical treatment.

[0975] SD 2, evening: rectal temperature: 40.5℃, overall health condition declined, taking Septomammin TM Topical treatment and use of Metacam TM (12 mL subcutaneous) treatment.

[0976] SD 3: Rectal temperature: 41°C, squeeze out the corresponding quadrant (yellowish fluid with teat sealant residue), use Orbenin Extra TM (1 intramammary applicator) treatment with Septomammin TM Topical treatment with Vetalgin TM (40 mL intravenously) treatment.

[0977] SD 4: Morning and evening: squeeze out the corresponding quadrants and use Orbenin Extra TM (1 intramammary applicator) treatment with Septomammin TM Topical treatment.

[0978] SD 5: Morning and evening: squeeze out the corresponding quadrants and use Orbenin Extra TM (1 intramammary applicator) treatment with Septomammin TM Topical treatment.

[0979] SD 6: general appearance decreased, ears drooping, animals often lying down, poor rumen filling, loss of appetite, swollen and hardened udder quadrants, no fever, pain on palpation, increased heart rate, rectal examination: no feces, moderate rumen filling, infusion therapy (1L of G40 TM , 60mL Catosal TM , 13mL Metacam TMintravenous), oral administration (40L water + 2 bags of Rumentinol TM Oral), squeeze out the corresponding quadrant (a small amount of yellowish fluid and teat sealant residue), Orbenin Extra TM (1 intramammary applicator).

[0980] SD 7: Rectal temperature: 37.8℃, oral administration (40L + 2 bags of Rumentinol TM ), the quadrant swelling was still severe, and the swelling moved to the right anterior nipple quadrant, and the patient was fed after the gavage.

[0981] SD 8: The right posterior quadrant remains firm and swollen, the swelling moves to the right anterior teat quadrant, the animal is healthier, the rumen is moderately full, mucous nasal discharge, infusion therapy (1.5L NaCl, 0.5L G40 TM ,30mL Catosal TM ,250mLCalciumborogluconat TM ,13mL Metacam TM ,5mL Mederantil TM intravenous), with a shaky gait after infusion but good feed intake.

[0982] SD 9: Overall appearance improved, feeding and rumination good.

[0983] SD 14: Overall appearance was good, with the right posterior quadrant still severely firm and swollen, and the right anterior quadrant firm and swollen.

[0984] Breast autopsy

[0985] Through deep anesthesia (with ketamine / xylazine) and subsequent T61 TM All animals were euthanized. Only the udder and mammary lymph nodes were collected and examined. Body weights were recorded before the start of the study.

[0986] After euthanasia, various samples were collected from each udder quadrant of each cow, e.g. Figure 67 shown.

[0987] In addition, samples from the area of ​​macroscopic findings and two mammary lymph nodes were collected from each cow.

[0988] Necropsy findings revealed areas in the left udder of one cow in the TS1 group. Changes in contents and consistency were observed in the right and left udders of one animal in the TS2 group. Mammary gland swelling was observed in one cow in the TS1 group. Umbilical lymphadenopathy was observed in two cows in the TS1 group and in one cow in the TS2 group.

[0989] Histopathology

[0990] For TS1, TS2 or Orbeseal TM No treatment-related local toxic effects were observed. Three cows were diagnosed with chronic active suppurative or subacute abscessive mastitis. Many cows had increased granulocytes and / or mononuclear cells or debris in one or more udder quadrants.

[0991] TS1 and 2 or Orbeseal TM A single intramammary administration of did not reveal any local toxic effects associated with the test substance. After two weeks, all cows were still clearly lactating. Despite this, two cows (I 360 and I 366) showed chronic active inflammation in one or both udder quadrants, which cannot be ruled out to have occurred before treatment and / or aggravated by dry milk. Reactive cells increased in the associated lymph nodes. Cow I 363 developed a subacute abscess in one quadrant, which was consistent with the clinical findings during treatment. Many cows did not have major histopathological findings, but had an increase in granulocytes and / or mononuclear cells or debris in the mammary alveoli, suggesting a udder degeneration process.

[0992] Mastitis was detected in 3 cows:

[0993] I 360 (Group 01: TS1 or Orbeseal TM ) had moderate chronic active suppurative inflammation in the right anterior quadrant and nipple (with Orbeseal TM In the case of a breast cancer treated with TS1, there was mild focal fibrosis in the glandular tissue. Correspondingly, the right mammary lymph node was grossly enlarged and slightly hypercellular microscopically. Mild focal glandular fibrosis was also observed in the right posterior and left anterior quadrants (both treated with TS1).

[0994] I 363 (Group 01: TS1 or Orbeseal TM ) revealed multiple subacute abscesses of significant severity in the right posterior quadrant with moderate bacterial counts (with Orbeseal TM Treatment). The nipple was affected by moderate inflammation. The right mammary lymph node showed slightly increased cellularity.

[0995] I 366 (Group 02: TS2 or Orbeseal TM ) affected by the left anterior quadrant (with Orbeseal TM Mild to moderate chronic active suppurative inflammatory involvement of the mammary gland and nipple in the left posterior quadrant (treated with TS2) and left posterior quadrant (treated with TS2) with focal glandular fibrosis. The associated left lymph node showed mild increased cellularity and lymphoid hyperplasia.

[0996] Noteworthy findings

[0997] Notable findings believed to be related to the test substances are summarized below in Table 8H.

[0998] Table 8H. Summary of noteworthy findings given at the lowest dose level.

[0999]

[1000] M+F = findings seen in both sexes at the same dose level

[1001] M / F = findings seen in both sexes at different dose levels

[1002] summary

[1003] Nipple Sealants 1 and 2 or Orbeseal within two weeks TM A single intramammary administration of did not reveal any local toxic effects related to the test substance. All cows continued to lactate significantly. Nevertheless, two cows (I 360 and I 366) showed chronic active inflammation in one or both udder quadrants, which cannot be excluded as occurring before treatment and / or aggravated by dry milk. Reactive cell counts were seen in the associated lymph nodes. Cow I 363 developed a subacute abscess in one quadrant, which was consistent with the clinical findings during treatment. Although there were no major histopathological findings in many cows, there was an increase in granulocytes and / or mononuclear cells or debris in the mammary alveoli, which may be the cause of the clinically increased cell counts in the milk.

[1004] in conclusion

[1005] The two adverse events of clinical mastitis that occurred after the dry period were unlikely to be related to IVP (i.e., TS1 and TS2). Both affected quadrants were treated with Orbeseal at SD 0. TM Sealants. Aside from milk dripping in three cows after the dry period, no further local tolerability findings were observed during the evaluation. Histopathological examination did not reveal any IVP-related local toxic effects. Both Teat Sealants 1 and 2 were well tolerated both systemically and locally.

[1006] Advantages of the invention as listed

[1007] Some advantages of certain embodiments of the invention cited include:

[1008] - The teat sealant formulation does not contain bismuth salts and has antimicrobial activity (but "antibiotic-free" means it does not contain antibiotics used in dry cow therapy);

[1009] -Teat sealant formulations are easy to apply;

[1010] -Nipple sealant preparations can be infused with a syringe;

[1011] -Teat sealant formulations will remain in the teat for most of the dry period;

[1012] -Nipple sealant formulations are easy to remove without the worry of leftover heavy metal residues, such as bismuth residues;

[1013] - At the end of the dry period, the teat sealant preparation can be squeezed out;

[1014] -Teat sealant formulations are effective and safe for cows; and

[1015] -Tit sealant preparations can be used to prevent mastitis.

[1016] In this specification, the word "comprise" and its derivatives (including "comprising" and "containing") include each stated integer but do not exclude the inclusion of one or more other integers.

[1017] The terms "about" and "approximately" represent an interval of accuracy that one skilled in the art will understand still ensures the technical effect of the feature in question. The term generally indicates a deviation of ±10%, preferably ±5%, more preferably ±2%, and even more preferably ±1% from the numerical value indicated.

[1018] Reference in this specification to any numerical range includes all possible numbers / values ​​within that range and further includes all possible sub-ranges within that range (if the context permits).

[1019] Reference in this specification to any previous publication (or information obtained therefrom) or any known matter does not constitute and should not be taken as an acknowledgment or endorsement or any form of implication that the previous publication (or information obtained therefrom) or known matter forms part of the common general knowledge in the field to which this specification relates.

[1020] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases "in one embodiment" or "in an embodiment" throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more combinations.

[1021] In accordance with the statute, the present invention has been described using language that is more or less specific to structural or methodological features. It should be understood that the present invention is not limited to the specific features shown or described, as the means herein described comprise preferred forms of carrying out the invention. Accordingly, the present invention is claimed in any form or modification within the proper scope of the appended claims as appropriately interpreted by one skilled in the art.

Claims

1. A teat sealant composition free of bismuth salts, formulated for occluding teat ducts and / or teat cistern.

2. A teat sealant composition comprising a salt other than a bismuth salt, formulated for occluding the teat duct and / or teat cistern.

3. A teat sealant composition formulated for application to and retention within the teat canal and / or teat cistern of a cow.

4. The teat sealant composition of any one of the preceding claims, wherein the teat sealant composition is in the form of a gel, a paste or a coarse suspension.

5. The teat sealant composition of any one of the preceding claims, wherein: - the teat sealant composition has a sufficiently low viscosity to allow it to be administered by injection through the teat canal and / or teat cistern; - Apply / inject approximately 2-5g of the teat sealant composition into the teat canal and / or teat cistern; - the injection force for injecting the teat sealant composition into the teat at 25°C does not exceed 4500g, and preferably the injection force for injecting the teat sealant composition into the teat at 25°C is between about 1500g and 3500g; - the teat sealant composition rapidly thickens in the teat, thereby being retained and blocking the teat ducts and / or teat cistern; - once located within the teat canal and / or teat cistern, the teat sealant composition has a sufficiently high viscosity to remain within and block the teat canal or teat cistern; - the teat sealant composition has a rebound viscosity enabling it to be retained within the teat canal and / or teat cistern as a substantially cohesive mass; The teat sealant composition has rheological properties that enable the composition to be administered into the teat duct and / or teat cistern by injection and to be retained in the teat duct and / or teat cistern to block the teat duct and / or teat cistern or to prevent intramammary infection; - The teat sealant composition has a viscosity greater than about 1 g / cm 3 The density is preferably about 1.3 g / cm 3 Up to 1.6g / cm 3 between; - the teat sealant composition shear thins under the shear forces applied during injection and milking from the teat; - the teat sealant composition has antimicrobial activity, preferably antibacterial activity, preferably against bacteria that cause mastitis; - the teat sealant composition is formulated to occlude the teat duct and / or teat cistern for a period of at least about 14 days, preferably about 42 to about 90 days; - the teat sealant composition is formulated to occlude the teat duct and / or teat cistern throughout the dry period; - the teat sealant composition is formulated to occlude the teat ducts and / or teat cisterns for at least about the first 14 days of the dry period until a natural keratin plug forms; - the teat sealant composition occludes or substantially occludes the teat ducts and / or teat cistern for about 4-6 weeks while allowing the natural keratin plug to form; or - The teat sealant composition is formulated so that it does not need to be milked from the teat canal and / or teat cistern at the end of the dry period.

6. A teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle, wherein the solid particles are not a bismuth salt.

7. The nipple sealant composition of claim 6, wherein: The antimicrobial activity is provided by the vehicle; the antimicrobial activity is provided by the solid particles; the antimicrobial activity is provided by both the vehicle and the solid particles; the antimicrobial activity is provided by a component of the vehicle; or, the composition further comprises at least one type of antimicrobial agent, preferably an antibacterial agent.

8. A teat sealant composition as claimed in claim 6 or claim 7, wherein the vehicle comprises a water insoluble carrier and a thickener, and preferably the thickener comprises the solid particles.

9. The nipple sealant composition of claim 8, wherein the water-insoluble carrier comprises at least one type of carrier agent, and the carrier agent comprises at least one type of oil or oily liquid.

10. The nipple sealant composition of claim 9, wherein the at least one type of oil or oily liquid comprises vegetable oil, mineral oil, synthetic oil, medium chain triglycerides or triglycerides, preferably paraffin oil, liquid paraffin, petrolatum, sesame oil or medium chain triglycerides.

11. The teat sealant composition of claim 9 or 10, wherein the composition comprises approximately 20% to 80% w / w or 30-65% w / w of at least one type of carrier agent.

12. The teat sealant composition of any one of claims 8 to 11, wherein the thickener comprises at least one type of water-soluble and / or water-insoluble thickening agent, preferably in an amount of about 0.1% to 10% w / w.

13. The teat sealant composition of claim 12, wherein the at least one thickening agent functions as a thickening agent and as an antimicrobial agent.

14. The teat sealant composition of claim 12 or claim 13, wherein the at least one thickening agent comprises a medium chain saturated fatty acid, preferably lauric acid.

15. The teat sealant composition of claim 14, wherein the composition comprises about 1% to 10% w / w medium chain saturated fatty acid, preferably lauric acid.

16. The nipple sealant composition of claim 12 or 13, wherein the at least one thickening agent comprises a metal salt of a long chain fatty acid, preferably a stearate.

17. The nipple sealant composition of claim 16, wherein the metal salt of a long-chain fatty acid comprises aluminum distearate, aluminum stearate, aluminum tristearate, ammonium stearate, barium stearate, butyl stearate, cadmium stearate, calcium stearate, cobalt stearate, copper stearate, ethylene glycol stearate, lithium stearate, magnesium stearate, manganese stearate, methyl stearate, potassium stearate, sodium stearate, strontium stearate, or zinc stearate, preferably aluminum stearate.

18. The teat sealant composition of claim 17, wherein the composition comprises about 1% to 10% w / w of the metal salt of a long chain fatty acid.

19. The nipple sealant composition of claim 12 or claim 13, wherein the at least one thickening agent comprises silica, preferably hydrophilic or hydrophobic silica.

20. The nipple sealant composition of claim 19, wherein the composition comprises about 0.1% to 1% w / w silica, preferably hydrophilic fumed silica.

21. The teat sealant composition of any one of claims 6 to 20, wherein: - the solid particles are very soluble, readily soluble, soluble, slightly soluble, slightly soluble, very slightly soluble, practically insoluble or insoluble in water; - the solubility of the solid particles in water exceeds about twice the isotonic concentration; - the solid particles modify the rheology of the composition as well as provide antimicrobial activity, preferably against mastitis-causing bacteria; - the solid particles comprise at least one type of salt other than a bismuth salt, such as an organic salt or an inorganic salt; - the solid particles comprise a halide salt, a phosphate or a carbonate, preferably sodium chloride, potassium chloride, calcium phosphate as defined herein, preferably calcium hydrogen phosphate dihydrate or magnesium carbonate; - the solid particles contain at least one type of heavy metal salt, provided that it is not a bismuth salt, preferably the metal of the heavy metal salt contains calcium, magnesium or aluminum, but does not include toxic heavy metals; - the solid particles comprise at least one type of carbohydrate, such as a simple sugar or a complex sugar, preferably sucrose or mannitol; - the solid particles comprise at least one type of mineral, such as clay, such as kaolin, such as kaolinite, halloysite, dickite or nacrite; - the solid particles contain sodium chloride; - the solid particles contain potassium chloride; - the solid particles comprise calcium phosphate as defined herein; - the solid particles comprise calcium hydrogen phosphate dihydrate; - the solid particles comprise calcium carbonate; - the solid particles comprise magnesium carbonate; - the solid particles comprise sodium chloride or potassium chloride and calcium phosphate as defined herein, or calcium hydrogen phosphate dihydrate; - the solid particles contain sodium chloride or potassium chloride and calcium carbonate; - the solid particles contain sodium chloride or potassium chloride and magnesium carbonate; - the composition comprises solid particles in an approximate amount ranging from 10% to 75%, 35% to 70%, 40% to 65% or 50% to 60% w / w; - the composition comprises 25-60% w / w sodium chloride or potassium chloride; - the composition comprises 10-30% w / w magnesium carbonate; - the composition comprises 10% w / w magnesium carbonate and approximately 25% w / w sodium chloride or potassium chloride; - the composition comprises 55% w / w sodium chloride or potassium chloride; - the composition comprises 35-60% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate; - the composition comprises 60% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate; - the composition comprises 10% w / w sodium chloride or potassium chloride and 50% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate; - the composition comprises 25% w / w sodium chloride or potassium chloride and 35% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate; or - the composition comprises 10% w / w sodium chloride or potassium chloride and 50% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate.

22. The teat sealant composition of any one of claims 6 to 21, further comprising one or more other ingredients such as at least one antioxidant or preservative.

23. A teat sealant composition comprising solid particles in the form of a gel or a paste or in the form of a coarse suspension formulated for occluding teat ducts and / or teat cistern.

24. A teat sealant composition comprising: an oily carrier; and a thickener comprising solid particles and lauric acid, wherein the solid particles are not bismuth salts.

25. A teat sealant composition comprising a coarse filler material in the form of solid particles in a gelling agent, wherein the solid particles are not a bismuth salt.

26. A teat sealant composition comprising: Liquid paraffin; Aluminum stearate; fumed silica; and Water-soluble solid particles, The composition is in the form of a paste or a suspension.

27. A teat sealant composition comprising: Liquid paraffin; Aluminum stearate; fumed silica; and Water-insoluble solid particles, The composition is in the form of a paste or a suspension.

28. The nipple sealant composition of any one of claims 23 to 27, wherein the average solid particle size is less than about 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80 μm in size, or the average solid particle size is about 10-50 μm in size, or the average solid particle size is about 1-10 μm in size.

29. The nipple sealant composition of any one of claims 23 to 28, wherein: - the solid particles are very soluble, readily soluble, soluble, slightly soluble, slightly soluble, very slightly soluble, practically insoluble or insoluble in water; - the solubility of the solid particles in water exceeds about twice the isotonic concentration; - the solid particles modify the rheology of the composition as well as provide antimicrobial activity, preferably against mastitis-causing bacteria; - the solid particles comprise at least one type of salt other than a bismuth salt, such as an organic salt or an inorganic salt; - the solid particles comprise a halide salt, a phosphate or a carbonate, preferably sodium chloride, potassium chloride, a calcium phosphate as defined herein, such as calcium hydrogen phosphate dihydrate or magnesium carbonate; - the solid particles contain at least one type of heavy metal salt, provided that it is not a bismuth salt, preferably the metal of the heavy metal salt contains calcium, magnesium or aluminum, but does not include toxic heavy metals; - the solid particles comprise at least one type of carbohydrate, such as a simple sugar or a complex sugar, preferably sucrose or mannitol; - the solid particles comprise at least one type of mineral, such as clay, such as kaolin, such as kaolinite, halloysite, dickite or nacrite; - the solid particles contain sodium chloride; - the solid particles contain potassium chloride; - the solid particles comprise calcium phosphate as defined herein; - the solid particles comprise calcium hydrogen phosphate dihydrate; - the solid particles comprise calcium carbonate; - the solid particles comprise magnesium carbonate; - the solid particles comprise sodium chloride or potassium chloride and calcium phosphate as defined herein, such as calcium hydrogen phosphate dihydrate; - the solid particles contain sodium chloride or potassium chloride and calcium carbonate; - the solid particles contain sodium chloride or potassium chloride and magnesium carbonate; - the composition comprises solid particles in an approximate amount ranging from 10% to 75%, 35% to 70%, 40% to 65% or 50% to 60% w / w; - the composition comprises 25-60% w / w sodium chloride or potassium chloride; - the composition comprises 10-30% w / w magnesium carbonate; - the composition comprises 10% w / w magnesium carbonate and approximately 25% w / w sodium chloride or potassium chloride; - the composition comprises 55% w / w sodium chloride or potassium chloride; - the composition comprises 35-60% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate; - the composition comprises 60% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate; - the composition comprises 10% w / w sodium chloride or potassium chloride and 50% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate; - the composition comprises 25% w / w sodium chloride or potassium chloride and 35% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate; or - the composition comprises 10% w / w sodium chloride or potassium chloride and 50% w / w calcium phosphate as defined herein, such as dibasic calcium phosphate dihydrate.

30. A teat sealant composition comprising one of the following formulations, wherein all ingredients shown are actual or approximate values: NaCl or KCl 55% w / w; Aluminum stearate 4.3% w / w; Fumed silica 0.7% w / w; Lauric acid 5% w / w; and Paraffin oil 35% w / w; or NaCl or KCl 60% w / w; Aluminum stearate 4.3% w / w; Fumed silica 0.7% w / w; Lauric acid 5% w / w; and Paraffin oil 30% w / w; or NaCl or KCl 55% w / w; Aluminum stearate 4.3% w / w; Fumed silica 0.7% w / w; Lauric acid 2.5% w / w; and Paraffin oil 37.5% w / w; or NaCl or KCl 10% w / w; Calcium phosphate as defined herein, such as calcium hydrogen phosphate dihydrate 50% w / w; Aluminium stearate 4.3% w / w; Fumed silica 0.7% w / w; Lauric acid 5% w / w; and Paraffin oil 30% w / w; or NaCl or KCl 25% w / w; Calcium phosphate as defined herein, such as calcium hydrogen phosphate dihydrate 35% w / w; Aluminium stearate 4.3% w / w; Fumed silica 0.7% w / w; Lauric acid 5% w / w; and Paraffin oil 30% w / w; or Calcium phosphate as defined herein, such as calcium hydrogen phosphate dihydrate 60% w / w; Aluminium stearate 4.3% w / w; Fumed silica 0.7% w / w; Lauric acid 5% w / w; and Paraffin oil 30% w / w; or Magnesium carbonate 25% w / w; Aluminum stearate 4.3% w / w; Fumed silica 0.7% w / w; Lauric acid 5% w / w; and Paraffin oil 65% w / w; or NaCl or KCl 50% w / w; Aluminum stearate 5.3% w / w; Fumed silica 0.7% w / w; Lauric acid 5% w / w; and Paraffin oil 39% w / w; or Magnesium carbonate 30% w / w; Fumed silica 0.7% w / w; Lauric acid 5% w / w; and Paraffin oil 64.3% w / w; or NaCl or KCl 50% w / w; Aluminum stearate 5.3% w / w; Fumed silica 0.7% w / w; and Paraffin oil 44% w / w; or NaCl or KCl 25% w / w; Magnesium carbonate 10% w / w; Aluminum stearate 4.3% w / w; Fumed silica 0.7% w / w; and Paraffin oil 60% w / w.

31. A method of preparing a teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle, the method comprising the steps of: (1) forming a water-insoluble gel by mixing at least one type of water-insoluble carrier agent with at least one type of thickening agent; (2) adding solid particles to the gel to form a paste; and, optionally, (3) adding at least one other type of thickening agent to the gel or paste, wherein: preparing a teat sealant composition comprising solid particles dispersed in a water-insoluble, shear-thinning, viscous fluid vehicle; The solid particles are not bismuth metal salts; Step (3) is optional; and Steps (2) and (3) do not need to be performed in the order described.

32. A method of preparing a teat sealant composition, the method comprising the steps of: (1) forming a water-insoluble gel by mixing at least one type of oily vehicle with at least one type of thickening agent; (2) adding solid particles to the gel to form a paste; and, (3) adding lauric acid as a thickening agent to the gel or paste, wherein: The solid particles are not bismuth metal salts.

33. A teat sealant composition prepared by the method of claim 31 or claim 32.

34. A teat sealant composition for preventing new intramammary infection or mastitis in a cow, wherein the composition is as defined in any one of claims 1 to 30 and 33.

35. A teat sealant composition for use in preventing new intramammary infection or mastitis in a cow or when used for such use, wherein the composition is as defined in any one of claims 1 to 30 and 33.

36. Use of a teat sealant composition in the preparation of a medicament for preventing new intramammary infection or mastitis in a cow, wherein the composition is as defined in any one of claims 1 to 30 and 33.

37. A method of preventing new intramammary infection or mastitis in a cow, said method comprising the step of applying a composition as defined in any one of claims 1 to 30 and 33 to at least one teat of the cow.

38. Use of a composition as defined in any one of claims 1 to 30 and 33 for preventing new intramammary infections or mastitis in cows.

39. A syringe containing a composition as defined in any one of claims 1 to 30 and 33.

40. A kit for use in a method of preventing a new intramammary infection or mastitis in a cow, or when used for such use, wherein the kit comprises a syringe capable of administering a composition as defined in any one of claims 1 to 30 and 33 to at least one teat of a cow.

41. The teat sealant composition of claim 34 or 35, the use of claim 36 or 38, the method of claim 37, or the kit of claim 40, wherein the composition is used in a dairy cow or heifer.

42. The nipple sealant composition of claim 34, 35 or 41, the use of claim 36, 38 or 41, the method of claim 37 or 41 or the kit of claim 40 or 41, wherein: - the composition prevents new intramammary infection or mastitis for a period of at least about 14 days, about 42 to about 90 days, or up to about 4 months; - the occlusion achieved by the composition is for at least the first approximately 14 days of the dry period, until the natural keratin plug is formed; - the composition occludes the teat duct and / or teat cistern or substantially occludes the teat duct and / or teat cistern until a natural keratin plug has formed in the teat duct; - the composition protects the cow's teat(s) from bacterial infection throughout the dry period; - administering the composition to each quadrant at the end of lactation; - administering the composition per quadrant in primiparous heifers approximately 4 weeks prior to calving; - the teat sealant composition has a sufficiently low viscosity to allow it to be administered by injection through the teat canal and / or teat cistern; - Apply / inject approximately 2-5g of the teat sealant composition into the teat canal and / or teat cistern; - the injection force for injecting the teat sealant composition into the teat at 25°C does not exceed 4500g, and preferably the injection force for injecting the teat sealant composition into the teat at 25°C is between about 1500g and 3500g; - the teat sealant composition rapidly thickens in the teat, thereby being retained in and blocking the teat ducts and / or teat cistern; - once located within the teat canal and / or teat cistern, the teat sealant composition has a sufficiently high viscosity to be retained within and block the teat canal or teat cistern; - the teat sealant composition has a rebound viscosity enabling it to be retained within the teat canal and / or teat cistern as a substantially cohesive mass; The teat sealant composition has rheological properties that enable the composition to be administered into the teat duct and / or teat cistern by injection and to be retained in the teat duct and / or teat cistern to block the teat duct and / or teat cistern or to prevent intramammary infection; - the teat sealant composition shear thins under the shear forces applied during injection and expression from the teat; or - The teat sealant composition occludes or substantially occludes the teat ducts and / or teat cistern for about 4-6 weeks while allowing the natural keratin plug to form.

Images