Fatty acid composition

Abstract

The present invention relates to a composition comprising a mixture of fatty acids, such as linoleic acid, oleic acid, and palmitic acid or derivatives thereof, derived from the secretions of the mammary gland of a mammal, which composition can be used to reduce stress, anxiety, and aggression in a mammal.

Description

[Technical field]

[0001] cross reference This application claims priority from, and is a continuation-in-part of, U.S. Patent Application No. 63 / 319,545, filed March 14, 2022, which is incorporated by reference herein.

[0002] The present invention relates to a composition comprising a mixture of fatty acids or derivatives solubilized in mineral oil, which composition is used to reduce the incidence of economically important diseases in livestock. [Background technology]

[0003] Stress is the negative response of the animal body to natural harmful forces, infections and various abnormal conditions that tend to disturb homeostasis.

[0004] Animals exposed to stress respond with changes in the activity of the autonomic nervous system and neuroendocrine system, as well as in behavior. The activation of these biological systems is a prerequisite for animals to cope with stress and is therefore the main resource for providing adequate biological defense against threats to the animal's homeostasis.

[0005] In animals, including humans, stress stimulates the release of adrenocorticotropic hormone (ACTH), which controls the release of cortisol from the adrenal cortex.

[0006] In humans, stress can cause medical problems such as ulcers, erosions, acute gastritis, and diarrhea. The development of polycythemia, inflammatory bowel disease, heart attacks, and ischemia are influenced by stress. There is strong clinical evidence that mental or emotional stress and anxiety are associated with the promotion of overt ischemic heart disease and sudden death.

[0007] Mammals such as pigs, dogs, and cows are also affected by changes in their environment. Removal of animals from their environment, herding together, and transportation often results in stressed animals. As a result, pathological disorders, death, growth retardation, and behavioral disorders often occur under stress-related conditions. Behavioral disorders often result in aggressive fighting when animals are stressed.

[0008] Social stress is common during development and often occurs as a result of separation from the dam, movement to a new environment, and mixing with unfamiliar juveniles.

[0009] This social stress often leads to antagonistic behavior between the animals, consisting of fighting or attempts to avoid each other. For example, piglets begin fighting within hours of birth. When pigs of any age meet, fighting is likely to occur. It has been suggested that pheromones are released towards the end of a fight that signal surrender.

[0010] In dairy cows, the transition period is defined as the period from 21 days before to 21 days after calving, which is known to be a critically challenging period in the dairy cow's life cycle. High-producing dairy cows undergo extreme metabolic adaptations during this period. Feed consumption increases approximately two-fold between the week before calving and the first 30 days after calving; however, nutrient intake is often insufficient to meet lactation demands. As a result of this catabolic state, high-producing dairy cows may lose 1.5 kg / day of body weight within the first 3 weeks after calving, when in energy balance with a net energy of lactation (NEL) of -7 to -9 Mcal / day. In addition, the transition period represents a tremendous accumulation of stressors for dairy cows. Pre-calving cows are transferred to individual calving pens during stage 1 of calving. Their newborn calves are separated from their dams immediately after calving. Post-calving cows are then transferred to post-calving lactation pens from where they are sent to the milking parlor three times a day. For first lactation cows, this is their first exposure to the milking parlor and milking procedures. Thus, during the transition period, the animals are exposed to incredible levels of social, nutritional, environmental, and physiological stress that often cause morbidity, mortality, and reduced productivity. Retained placenta, metritis, mastitis, and lameness are among some of the diseases that afflict post-calving dairy cows.

[0011] Retained fetal membranes (RFM) is defined as the failure to expel the placenta within 24 hours after delivery. It has been hypothesized that RFM may result from impaired immune function during the perinatal period, with a particular focus on neutrophil function and migration. Thus, failure to abruptly release the placenta appears to be due, at least in part, to a reduced ability of neutrophils to digest the cotyledon-caruncle attachment after delivery.

[0012] Metritis is defined as an inflammatory process of the uterus occurring within 21 days after calving and characterized by a foul-smelling, reddish-brown, watery uterine vaginal discharge accompanied by signs of systemic illness and a rectal temperature (RT) >39.5°C. If no systemic signs are present and RT <39.5, the condition can be defined as clinical metritis. Metritis is associated with reduced milk production, poor reproductive performance, high treatment costs and increased risk of culling, resulting in significant economic losses.

[0013] Mastitis is a highly prevalent disease in dairy cows and arguably the most important disease for the dairy industry worldwide, causing economic losses due to reduced milk production, milk discards, lower conception rates, premature culling, and treatment costs. The well-documented loss of milk production due to mastitis is estimated at approximately 15% of the milk-producing potential of affected cows. Clinical mastitis is also a serious animal welfare challenge, as it is associated with pain, poor health, and behavioral changes.

[0014] Despite decades of research contributing to advanced knowledge and support in the prevention of perinatal disorders (e.g., lameness, mastitis, retained placenta, and metritis), their incidence remains high. Moreover, these infections are often treated with systemic or local antibiotic therapy. The emergence of antibiotic-resistant bacteria is a growing concern, as many of the antibiotics used in animal husbandry are also available in human medicine. In light of this, there is a growing demand for alternatives to antibiotic use and more effective treatment options that could improve the health of dairy cows.

[0015] Diarrhea is one of the leading causes of mortality and economic losses during the preweaning period of domestic mammals. It is also one of the most common (18.2%) diseases in preweaned young cows. Calves during the first month of life have the greatest risk of developing diarrhea. Furthermore, the incidence of diarrhea in young calves has been found to be a causative agent for other diseases later in life, such as pneumonia. Diarrhea can develop as a result when newborn calves are exposed to various infectious or non-infectious risk factors. The most important risk factors in newborn calves are pathogen exposure, such as nutritional management, environmental conditions, and hygiene. The most common enteric pathogens associated with diarrhea in the first month of life in calves are rotavirus, coronavirus, E. Coli, Cryptosporidium spp., and coccidia.

[0016] Pageat (U.S. Pat. No. 6,169,113) (hereinafter, the "'113 patent") discloses a pheromone composition (see Table 1) that may reduce stress, anxiety, and aggression in mammals. The claimed pheromone formulation is described in claim 2 of the '113 patent as "a method for reducing mortality and morbidity during infection in a mammal, comprising administering to a mammal in need of such treatment a solution containing, as primary active ingredients, palmitic acid, oleic acid, linoleic acid and derivatives thereof, said solution comprising about 18%-31.2% (w% / w%) palmitic acid, about 34.3%-47.2% (w% / w%) linoleic acid, and about 28.7%-42.8% (w% / w%) oleic acid and derivatives thereof," as well as in claim 3 of the '113 patent. The disclosure of claim 3 of the Patent provides, "A method for reducing mortality and morbidity during infection in a mammal, comprising administering to a mammal in need of such treatment a solution containing, as main active ingredients, palmitic acid, oleic acid, linoleic acid and derivatives thereof, said solution containing about 18% to 31.2% (w% / w%) palmitic acid, about 34.3% to 47.2% (w% / w%) linoleic acid, about 8.7% to 16% (w% / w%) palmitoleic acid, and about 15.7% to 30.7% (w% / w%) oleic acid and derivatives thereof."

[0017] Fatty acids (FA) were dissolved and homogenized in 94% propylene glycol and 6% absolute alcohol at 37.5°C. A study was disclosed to evaluate the pheromone effect on piglets. Piglets were grouped and tested using propylene glycol as placebo and a 5% concentration of the pheromone formulation as treatment. In summary, it was reported that piglets receiving the placebo were 86% more aggressive and stressed than piglets treated with the pheromone composition. Furthermore, the placebo group had a loss of body weight compared to the treatment group. However, no effect on disease prevention was reported.

[0018] [Table 1] [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 shows the effect of treatment with a novel fatty acid composition on serum cortisol levels following castration. [Diagram 2] 1 shows the effect of treatment with a novel fatty acid composition on serum substance P levels following castration. [Diagram 3] FIG. 1 shows the effect of treatment with a novel fatty acid composition on weight gain after castration. [Figure 4] FIG. 1 shows the effect of treatment with a novel fatty acid composition on serum cortisol levels following dehorning. [Diagram 5] FIG. 1 shows the effect of treatment with a novel fatty acid composition on serum substance P levels following dehorning. [Figure 6] FIG. 1 shows the effect of treatment with a novel fatty acid composition on weight gain after dehorning. Summary of the Invention

[0020] It is an object of the present invention to provide a treatment for the prevention or amelioration of economically important diseases in livestock.

[0021] Another object of the present invention is to provide a composition that enhances weight gain in a mammal.

[0022] Another object of the present invention is to reduce mortality and morbidity in mammals.

[0023] Another object of the present invention is to reduce the incidence of retained placenta in mammals.

[0024] Another object of the present invention is to reduce the incidence of metritis in mammals.

[0025] Another object of the present invention is to reduce the incidence of lameness in mammals.

[0026] Another object of the present invention is to reduce the incidence of mastitis in mammals.

[0027] Another object of the present invention is to reduce feed conversion efficiency; that is, the ratio of food consumed / body weight gained.

[0028] Another object of the present invention is to alleviate pain experienced by a mammal being subjected to a painful procedure (e.g., castration, dehorning, or accidental physical injury) comprising the step of administering a pharma- tically effective amount of a composition of the present invention to a mammal in need thereof.

[0029] Another object of the present invention is to reduce the stress experienced by a mammal being subjected to a painful procedure (e.g., castration, dehorning, or accidental physical injury) comprising the step of administering a pharma- tically effective amount of a composition of the present invention to a mammal in need thereof.

[0030] Another object of the present invention is to enhance weight gain in a mammal after undergoing a painful procedure (e.g., castration, dehorning, or accidental physical injury) comprising the step of administering a pharma- tically effective amount of a composition of the present invention to a mammal in need thereof.

[0031] In one composition embodiment, the present invention provides a composition of eight fatty acids dissolved in mineral oil.

[0032] In another composition aspect, the present invention provides a basic fatty acid composition comprising palmitic acid, oleic acid, palmitoleic acid, linoleic acid, and derivatives thereof, which also has the ability to reduce the incidence of disease in livestock.

[0033] In another composition aspect, the present invention provides a composition comprising linoleic acid, palmitic acid, palmitoleic acid, oleic acid, arachidonic acid, myristic acid, lauric acid, capric acid, and derivatives thereof, dissolved in mineral oil. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] In the present invention, a new formulation of eight FAs is presented, which contains arachidonic acid, a major member of the omega-6 series, whose derivatives include endocannabinoids, which are oxidation-independent derivatives of critical significance for brain reward signaling, motivational processes, emotion, stress response, pain, and energy balance.

[0035] This fatty acid was not listed in the '113 patent or any other publication for this purpose. Furthermore, the percentage of FA was redefined. The FA of the present formulation is shown in Table 2.

[0036] More importantly, the complete fatty acid formulation is effective in dramatically reducing the incidence of several economically important diseases such as diarrhea, retained placenta, metritis, mastitis, and lameness. Mortality is also reduced in animals treated with the novel fatty acid formulation. Reduction in pain and pain-related stress have also been observed in mammals treated with the fatty acid formulation of the present invention. These effects are novel and were not listed in the '113 patent or any other publications for this purpose.

[0037] [Table 2]

[0038] Unlike propylene glycol, which is the solvent listed in the '113 patent, the solvent of the present composition is mineral oil. As used herein, the term "mineral oil" is any of a variety of colorless, odorless, light mixtures of higher alkanes from mineral sources, especially distillates of petroleum. Mineral oil is widely used as a moisturizing agent to treat or prevent dry, rough, scaly, itchy eczema and mild dermatitis, which is an important feature of the present invention since the application of the pheromone is topical. Mineral oil is a colorless, clear, oily liquid that is odorless and tasteless and is considered GRAS (Generally Recognized As Safe). Pharmaceutical grade mineral oil, food grade mineral oil, and / or industrial grade mineral oil are all liquid by-products of refining crude oil to make gasoline and other petroleum products. Mineral oil is a clear, colorless oil, related to petrolatum, composed primarily of alkanes and cycloalkanes. It has a density of about 0.8-0.87 g / cm 3 (0.029-0.031 lb / cu in). All grades of mineral oil are useful in the formulations of the present invention.

[0039] One embodiment of the present invention is a basic fatty acid composition comprising about 0.5% to 42.8% (wt% / wt%) of oleic acid and its derivatives.

[0040] Another embodiment of the present invention is a second basic fatty acid composition comprising about 0.5%-52.2% (wt% / wt%) of myristic acid and derivatives thereof.

[0041] Another embodiment of the present invention is a third basic fatty acid composition comprising about 0.5%-42.8% (wt% / wt%) oleic acid and 0.5%-52.2% (wt% / wt%) myristic acid and derivatives thereof.

[0042] Another embodiment of the present invention is a fourth basic fatty acid composition comprising about 0.5%-42.8% (wt% / wt%) oleic acid, 0.5%-52.2% (wt% / wt%) myristic acid, and 3%-31.2% (wt% / wt%) palmitic acid, and derivatives thereof.

[0043] Another embodiment of the present invention is a fifth basic fatty acid composition comprising approximately 0.5%-42.8% (wt% / wt%) oleic acid, 0.5%-52.2% (wt% / wt%) myristic acid, 3%-31.2% (wt% / wt%) palmitic acid, and 0.5-25% (wt% / wt%) arachidonic acid, and derivatives thereof.

[0044] Another embodiment of the present invention is a sixth basic fatty acid composition comprising approximately 0.5%-42.8% (wt% / wt%) oleic acid, 0.5%-52.2% (wt% / wt%) myristic acid, 3%-31.2% (wt% / wt%) palmitic acid, 0.5-25 (wt% / wt%) arachidonic acid, 0.2%-12.5% ​​(wt% / wt%) lauric acid, and derivatives thereof.

[0045] Another preferred embodiment of the present invention is a fatty acid composition comprising approximately 0.5%-42.8% (wt% / wt%) oleic acid, 0.5%-52.2% (wt% / wt%) myristic acid, 3%-31.2% (wt% / wt%) palmitic acid, 0.5-25% (wt% / wt%) arachidonic acid, 0.2%-12.5% ​​(wt% / wt%) lauric acid, 0.1%-22.6% (wt% / wt%) capric acid, and derivatives thereof.

[0046] Another preferred embodiment of the present invention is a fatty acid composition comprising approximately 0.5%-42.8% (wt% / wt%) oleic acid, 0.5%-52.2% (wt% / wt%) myristic acid, 3%-31.2% (wt% / wt%) palmitic acid, 0.5-25% (wt% / wt%) arachidonic acid, 0.2%-12.5% ​​(wt% / wt%) lauric acid, 0.1%-22.6% (wt% / wt%) capric acid, 8%-65% (wt% / wt%) linoleic acid, and derivatives thereof.

[0047] Another preferred embodiment of the present invention is a fatty acid composition comprising approximately 0.5%-42.8% (wt% / wt%) oleic acid, 0.5%-52.2% (wt% / wt%) myristic acid, 3%-31.2% (wt% / wt%) palmitic acid, 0.5-25% (wt% / wt%) arachidonic acid, 0.2%-12.5% ​​(wt% / wt%) lauric acid, 0.1%-22.6% (wt% / wt%) capric acid, 8%-65% (wt% / wt%) linoleic acid, 0.1%-12.6% (wt% / wt%) behenic acid, and derivatives thereof.

[0048] Another embodiment of the invention is a fatty acid mixture dissolved in mineral oil at a concentration ranging from 1% to 35% (wt% / wt%). Homogenization is easily achieved at concentrations as high as 35% by heating the mixture to 60° C. with vigorous stirring. Solutions are stable at concentrations as high as 35% without saturation or saponification.

[0049] Another embodiment of the invention is a method for treating stress, anxiety, aggressive behavior, or weight loss in a mammal, comprising the step of administering a pharma- tically effective amount of a composition of the invention to a mammal in need of such treatment.

[0050] Another embodiment of the invention is a method for reducing mortality and morbidity in a mammal, comprising the step of administering a pharma- tically effective amount of a composition of the invention to a mammal in need of such treatment.

[0051] Another embodiment of the invention is a method for reducing the incidence of diarrhea in a mammal, comprising administering to a mammal in need of such treatment a pharma- tically effective amount of a composition of the invention.

[0052] Another embodiment of the present invention is a method for reducing the incidence of retained placenta in a mammal, comprising the step of administering a pharma- tically effective amount of a composition of the present invention to a mammal in need of such treatment.

[0053] Another embodiment of the present invention is a method for reducing the incidence of metritis in a mammal, comprising the step of administering a pharma- tically effective amount of a composition of the present invention to a mammal in need of such treatment.

[0054] Another embodiment of the present invention is a method for reducing the incidence of mastitis in a mammal, comprising administering to a mammal in need of such treatment a pharma- tically effective amount of a composition of the present invention.

[0055] Another embodiment of the invention is a method for reducing the incidence of lameness in a mammal, comprising administering to a mammal in need of such treatment a pharma- tically effective amount of a composition of the invention.

[0056] Another embodiment of the present invention is a method for improving feed conversion in a mammal, comprising the step of administering a pharma- tically effective amount of a composition of the present invention to a mammal in need of such treatment.

[0057] Another embodiment of the invention is a method for reducing weight loss during transport comprising administering a pharma- tically effective amount of a composition of the invention to a mammal in need of such treatment.

[0058] Another embodiment of the present invention is a method for increasing carcass weight, comprising administering a pharma- tically effective amount of a composition of the present invention to a mammal in need of such treatment.

[0059] Another embodiment of the present invention is a method for alleviating pain experienced by a mammal undergoing a painful procedure, comprising the step of administering to a mammal in need of such treatment a pharma- ceutical effective amount of a composition of the present invention.

[0060] Another embodiment of the present invention is a method for reducing stress experienced by a mammal being exposed to a painful procedure, comprising the step of administering a pharma- tically effective amount of a composition of the present invention to a mammal in need of such treatment.

[0061] Another embodiment of the present invention is a method for enhancing weight gain in a mammal undergoing a painful procedure, comprising the step of administering to a mammal in need of such treatment a pharma- tically effective amount of a composition of the present invention.

[0062] As used herein, the term "mammals" includes any group of vertebrate animals, the females of which possess lactating glands. The term "mammals" includes, but is not limited to, cats, humans, dogs, pigs, rats, mice, cows, horses, apes, chimpanzees, dolphins, porpoises, killer whales, and the like.

[0063] "Stress" refers to the reaction of the animal body to harmful natural infectious forces and to various abnormal conditions that tend to disturb homeostasis. This reaction may be a physical reaction or an emotional reaction, such as anxiety.

[0064] "Anxiety" refers to a reactive state characterized by a high probability of fearful behavior and emotional responses, a feeling of anger and fear accompanied by restlessness, tension, etc. In neurophysical terms, this state of anxiety involves hyperactivity of the noradrenergic and serotonin systems.

[0065] "Aggressive behavior" refers to aggressive or defensive social interactions between individuals of the same species, such as fear, flight, or submission.

[0066] "Stress-related disorder" means any disorder whose symptoms increase due to stress.

[0067] "Reducing morbidity and mortality caused by an infection" means that mortality and morbidity are reduced so that the mammal has a better chance of survival.

[0068] "Improved feed conversion efficiency" means a reduction in the ratio of food intake / body weight gained.

[0069] The term "solution" means a liquid having one or more compounds dispersed therein, either by dissolution or suspension (eg, a colloidal suspension).

[0070] "Alleviating effect" means a reduction in fear, apprehension, or anxiety, as well as behavioral and physical effects associated with stress. Behavioral effects associated with stress include tremors, vocalizations, flight, aggression, dislocation behavior, etc. Physical effects associated with stress include changes in heart rate, levels of epinephrine, norepinephrine, ACTH, cortisol, glucose, etc. In animals used as a food source, this definition includes husbandry parameters such as growth weight and feed conversion efficiency.

[0071] "Basic fatty acid composition" means a fatty acid composition that is cross-species usable in all mammals and contains at least five fatty acids as the main active ingredients, such as oleic acid, myristic acid, palmitic acid, arachidonic acid, and lauric acid, and their derivatives. Optionally, the composition also contains one to four additional FAs, such as capric acid, palmitoleic acid, linoleic acid, behenic acid, and their derivatives.

[0072] "Enhancer composition" means an active pheromone composition that is species specific and is used to enhance or act synergistically with a basic pheromone composition to increase the effectiveness of the basic composition in a particular species.

[0073] More specifically, the basic compositions of the present invention comprise a mixture of at least five fatty acids (i.e., oleic acid, myristic acid, palmitic acid, arachidonic acid, and lauric acid) and their derivatives, representing about 15% to 95% (wt% / wt%) of the total composition, with the remaining components being mineral oils, and optionally other non-toxic filler compounds, such as fatty acids, alcohols, amines, squalene, and glycerol (more specifically, caproic acid, azelaic acid, propionic acid, geraniol, octadecatrianol, hexacosanol, trimethylamine, and methylamine).

[0074] The basic composition can also be attached to a chemical carrier, provided that the biologically active structure of the fatty acid is preserved. Such carrier molecules include, but are not limited to, resins, liposomes, crown compounds (e.g., macrocyclic multidentate), carrier proteins, and the like.

[0075] The fatty acids can be used in their pure form, i.e. as free fatty acids, as well as in the form of their derivatives, such as esters of fatty acids or salts of fatty acids, for example potassium laurate, potassium myristate, and potassium oleate.

[0076] The basic fatty acid composition can be diluted with a variety of solutions and can also be used in a variety of forms, as described below.

[0077] Enhancer compositions containing 5% to 35% (w% / w%) can also be added to the basic pheromone composition, if desired. The enhancer compositions include volatile organic compounds and mixtures thereof. The enhancer compositions may be species-specific in nature and may vary depending on the species selected for use in the present invention.

[0078] Compounds that may be used in the enhancer composition include, but are not limited to, amines and fatty acids from indole derivatives, esters of these amines and fatty acids, ketones, such as acetone, alcohols, sterols, and the like.

[0079] In addition to the basic fatty acid composition and the enhancer composition, a preferred embodiment of the present invention relates to a composition containing free fatty acids, including decanoic acid, such as capric acid, dodecanoic acid, such as lauric acid, tetradecanoic acid, such as myristic acid, hexadecanoic acid, such as palmitic acid, cis-9-octadecanoic acid, such as oleic acid, linoleic acid and palmitoleic acid. Derivatives of these fatty acids can also be used in the present invention. More specifically, these derivatives are fatty acid esters or fatty acid salts.

[0080] The compositions of the present invention may be in the form of a solution, an aerosol spray, a gel, a sustained release matrix, a shampoo, a microencapsulated product, etc. The solutions may also be used as topical treatments that are applied directly to the skin of an animal. More specifically, the basic fatty acid compositions may be applied as topical pour-on solutions to the skin of an animal.

[0081] The fatty acid concentration may vary depending on the final form of use. However, any specific fatty acid concentration that can be used may be used, and these concentrations can be confirmed and tested according to the methods described herein. In Example 3, adult Holstein dairy cows received a 1,000 milligram dose, which was applied topically to the skin on the neck and above the muzzle. The average live weight of adult Holstein dairy cows was 680 kg, and therefore the dose administered to the animals in Example 3 was 1.47 milligrams per kilogram of live weight. In Example 4, newborn dairy calves were treated with a 1,000 milligram dose, which was applied topically to the skin on the neck and above the muzzle. The average live weight of newborn Holstein calves is 45 kg. Thus, in Example 4, Holstein dairy calves received a dose of 22.22 milligrams per kilogram of live weight. In both examples, the administration of fatty acid concentrations dramatically reduced disease incidence and mortality. Thus, the effective dosage of the fatty acid composition may vary from 0.1 mg / kg to 1,000 mg / kg.

[0082] Once the compositions of the present invention are obtained, they can be tested for their effectiveness in treating stress and disease in mammals. Well-documented stressors include, for example, extreme cold, overcrowding, isolation, separation, confinement, closeness to predators, transportation, pain, etc. Application of the compositions in the form of sprays, aerosols, etc., in the area around the stress source results in a reduction in stress as indicated by various factors such as weight gain, social behavior with respect to other mammals, wounds on the body, especially the ears, salivary cortisol, and heart rate.

[0083] Thus, the composition can be applied to a variety of objects such as when a mammal is in close proximity to or in contact with walls, air, and toys, etc. The composition can also be applied to skin, hair, or fur.

[0084] In order to further illustrate the present invention, the following examples are given and it is understood that they are intended to be illustrative and not limiting. EXAMPLES

[0085] Example 1 Novel fatty acid composition The fatty acids used and their amounts are presented in Table 3. In total, eight different FAs were used. First, the four dry ingredients were weighed and transferred into a 2-liter glass bottle; 16 grams of palmitic acid, 10 grams of myristic acid, 8 grams of lauric acid, and 1 gram of behenic acid. Then, using a graduated pipette, the four liquid FAs were aliquoted and distributed into the same container; 30 mL of linoleic acid, 24 mL of oleic acid, 8 mL of arachidonic acid, and 3 mL of capric acid. To the bottle containing the FA mixture, 900 mL of mineral oil was added and the mixture was heated to 60° C. The final solution contained a concentration of 10% of the final fatty acid mixture and 90% of mineral oil. The solution was vigorously homogenized at 60° C. until complete dissolution of the crystals. Once the homogenized solution was obtained, the bottle was left to stand at room temperature and cooled. The resulting final product was sealed in a 30 mL glass vial and sealed at room temperature for long-term storage. The bottles were identified by batch number, date, and composition. Several batches were run as above to verify the product characteristics and stability. Visual inspection of the bottles was performed periodically and did not indicate any changes in the product. The product was stable, homogenous, and colorless.

[0086] [Table 3]

[0087] Example 2 Comparison of compositions Two fatty acid compositions were prepared and used to compare the formulations described in Table 3 using mineral oil as the solvent and using 94% propylene glycol and 6% ethanol (listed in the '113 patent) as the solvent. The fatty acid concentration of the compositions was 20% wt / wt.

[0088] Mineral oil with 94% propylene glycol and 6% ethanol were heated to 60°C. All FA was weighed and transferred to bottles to make 500mL compositions with 20% FA. After heating, each solvent was added to each bottle. The compositions were heated until complete FA dissolution and homogenized. The 20% FA mixture was used to make compositions with 1%, 5%, and 10% FA. The solvent was heated for proper dilution and homogenization. All flasks were well sealed and then stored at room temperature.

[0089] Visual inspection of the compositions was performed 24 and 72 hours after preparation. The experiment and visual inspection were repeated several times to verify the consistency of the solubility and stability of the solutions.

[0090] The compositions prepared with propylene glycol and ethanol were opaque and whitish, while the compositions prepared with mineral oil were colorless and translucent. After 24 hours, all compositions prepared with propylene glycol and ethanol were visually heterogeneous, opaque and whitish in the upper oil layer. On the other hand, the mineral oil compositions were stable, colorless, translucent and homogeneous. Visually, the solubility of the fatty acid compositions in mineral oil is substantially higher. After 72 hours, the compositions prepared with propylene glycol and ethanol containing 10% and 20% FA have visible precipitates and clumps of FA on the surface.

[0091] Absorbance (Ab) measurements were collected to more fully demonstrate the differences between the solvent and FA concentrations. The mineral oil compositions exhibited virtually the same Ab independent of FA concentration. However, the 94% propylene glycol and 6% ethanol compositions exhibited gradually increasing Ab measurements depending on the FA concentration. Using 1% FA, the mineral oil composition exhibited an Ab of 0.042, while propylene glycol and ethanol had an Ab of 0.197. When the 20% FA solution was measured, Abs of 0.059 and 1.385 were detected for mineral oil and propylene glycol and ethanol, respectively.

[0092] Example 3 Cow Test The study was conducted on Holstein cows on a dairy farm. All cows were fed a total mixed ratio (TMR) and the ration was pushed up eight times per day. The diet consisted of approximately 55% forage and 45% concentrate on a dry weight basis. The diet was formulated to meet or exceed the National Research Council nutrient requirements for a lactating Holstein cow weighing 650 kg and producing 45 kg of 3.5% fat corrected milk (National Research Council, 2001). Cows showing signs of parturition were transferred to individual maternity pens for parturition. After parturition, cows were transferred to a postpartum pen where they remained for approximately 40 days.

[0093] In total, 107 cows were enrolled in the study. Cows were divided into blocks by calving number (first calving and subsequent calvings) and randomly assigned within blocks to one of two treatment groups: control (CTR; n=53) and treatment (TRT; n=54). Cows were eligible to receive their respective treatments after they were detected to be showing signs of calving and were transferred to individual maternity pens for calving. In the maternity pens, cows assigned to the control group did not receive any treatment and cows assigned to the treatment groups received the following topical application of a novel fatty acid formulation: 5 mL applied to the skin on the neck and 5 mL applied to the skin above the muzzle. The fatty acid composition (10% fatty acid mixture and 90% mineral oil) was prepared as detailed in Example 1.

[0094] Retained fetal membranes were defined as not delivering fetal membranes by 24 hours postpartum. Metritis was diagnosed daily. Cows were flagged for physical examination when they showed signs of lethargy and depression or when >4.5 kg of milk abnormalities were detected. At physical examination, cows were considered to have metritis when a foul-smelling, watery, reddish-brown uterine vaginal discharge was noted in combination with systemic illness. Mastitis and lameness were also diagnosed by trained animal husbandry personnel.

[0095] Cows treated with the fatty acid formulation of the present invention had the following: Lower incidence of retained placenta (CTR=5.66% vs. TRT=1.84%) Lower incidence of metritis (CTR=15.1% vs. TRT=3.7%) Lower incidence of claudication (CTR=7.6% vs. TRT=3.7%) Lower incidence of mastitis (CTR=7.6% vs. TRT=5.6%).

[0096] Example 4 Dairy Calf Testing The objective of this study was to evaluate the effect of a mitigation pheromone composition of the present invention administered on the first day of birth and every 15 days thereafter on the incidence of diarrhea and risk of mortality in Holstein dairy calves.

[0097] The study was carried out on a dairy farm. Immediately after calving, calves were separated from their mothers and placed in neonatal pens lined with dry wood chips and heated by heat lamps at low temperatures for several months. All calves were fed approximately 4 L of pooled pasteurized colostrum by esophageal feeder (Oral Calf Feeder Bag with Probe, Jorvet) within 5 hours of birth. Once a day, calves were removed from the neonatal pens and transported to a calf rearing facility and placed in individual calf hutches. Calves were milked twice a day and one week after birth, solid feed was started. On the 50th day of calf birth, the weaning process was started by reducing the amount of milk offered, which continued until the 60th day of birth when milk was completely discontinued. After weaning, calves were kept in the hutches for approximately 2 weeks and then moved to group pens with a mixed population of 20 calves per pen.

[0098] The experimental design was a complete randomized design. A total of 420 Holstein dairy calves were randomly enrolled into one of two treatment groups (treatment, N=210; control, N=210). Calves in the treatment group received a single dose containing the novel fatty acid composition of the present invention (10 mL) as an initial treatment of the topical novel fatty acid formulation of the present invention (Example 1, Table 3) on their first day of birth, immediately after they were assigned to the calf hutch, and then every 14 days until weaning. The treatment was applied via topical administration to the skin on the neck (5 mL) and the skin above the muzzle (5 mL). Calves assigned to the control group did not receive any type of treatment. The novel fatty acid composition (10% fatty acid mixture and 90% mineral oil) was prepared as detailed in (Example 1, Table 3). All enrolled calves were evaluated daily over a 60-day follow-up period.

[0099] Statistical analyses were performed. Logistic regression was performed to analyze the effect of treatment on the binary response variables of incidence of diarrhea and risk of death. Fisher's exact significance test was used, considering the alternative hypothesis that the odds of diarrhea were higher in the control group compared to the treatment group.

[0100] Treatment reduced the incidence and risk of mortality of diarrhea during the preweaning period in calves. Administration of the novel fatty acid formulation significantly reduced (P<0.009) the incidence of diarrhea, which was diagnosed in 70.8% of calves assigned to the control group and 58.6% of calves assigned to the treatment group.

[0101] The risk of death within 60 days of birth was statistically higher in the control group (7.8%) compared with the treatment group (2.4%) (P=0.02).

[0102] Example 5 Beef Calf Testing Two studies were conducted to evaluate the effect of administering a novel fatty acid formulation on preventing weight loss after transportation. A novel fatty acid composition (10% fatty acid mixture and 90% mineral oil) was prepared as detailed in Example 1, Table 3. In studies 1 and 2, the novel fatty acid composition was administered to weaned beef calves prior to transport from the sale barn to the feedlot to evaluate weight loss during transport ("shrinkage").

[0103] Study 1 - The experiment was conducted with calves being transported from a receiving pen to a sales barn. A total of 38 calves, with an average weight of 272 kg, were used in the study. Calves were randomly assigned to either a treatment group (TRT, n=19) or a control group (CTR, n=19). Animals in the treatment group received a topical application of the novel fatty acid composition. Treatment was applied within 1 hour before loading onto the trailer. A total dose of 10 mL was applied topically to the skin on the nuchal (5 mL) and the skin above the muzzle (5 mL). Control animals did not receive any treatment and were loaded into a different trailer, avoiding contact with the treatment groups. The transport time from one of the animal facilities to the other was approximately 5 hours. The weight of the groups was measured before transport and upon arrival at the final destination.

[0104] After approximately 5 hours of transport, calves in the control group had lost a total of 415.5 kg (8%), whereas treated calves had lost a total of only 71.2 kg (1.3%). Thus, use of the novel fatty acid formulation prevented 83.75% of the weight loss observed in the untreated group.

[0105] Study 2 - The experiment was conducted with calves transported from sale barn to feedlot. The transport time of the animals from sale barn to feedlot was 12.5 hours. A total of 88 beef calves, with an average weight of 250 kg, were used in the study. Calves were randomly assigned to either a treatment group (TRT, n=19) or a control group (CTR, n=19). Animals in the treatment group received a topical application of a novel fatty acid composition, where a total dose of 10 mL was applied topically to the skin on the neck (5 mL) and the skin above the muzzle (5 mL). The novel fatty acid composition (10% fatty acid mixture and 90% mineral oil) was prepared as detailed in Example 1, Table 3. The treatment was applied within 1 hour before loading onto the trailer. The control animals did not receive any treatment and were loaded onto a different trailer, avoiding contact with the treatment group. The weight of the group was measured before transport and upon arrival at the final destination.

[0106] After 12.5 hours of transport, calves in the control group had lost a total of 902.6 kg (7.55%), while calves in the treatment group had lost a total of 13.6 kg (0.11%). Thus, use of the novel fatty acid formulation prevented 98.54% of the weight loss observed in the untreated group.

[0107] Example 5 Final beef cattle testing The experiment was carried out with fat beef cattle (finishing beef cattle) being transported from a feedlot to a slaughterhouse. A total of 76 beef steers with an average live weight of 583 kg were used in the study. Steers were randomly assigned to either a treatment group (TRT, n=38) or a control group (CTR, n=38). Animals in the treatment group received topical application of the novel fatty acid composition. Treatment was applied within 12 hours before loading onto the trailer. A total dose of 10 mL was applied topically to the skin on the neck (5 mL) and the skin above the muzzle (5 mL). Control animals did not receive any treatment and were loaded into a different trailer, avoiding contact with the treatment groups. The novel fatty acid composition (10% fatty acid mixture and 90% mineral oil) was prepared as detailed in Example 1, Table 3. The transport time of the animals from the feedlot to the slaughterhouse was approximately 5 hours. Individual live weights were recorded before transport and warm carcass weights (HCW) were recorded at the slaughterhouse.

[0108] Before transport, the study steers had a mean live weight of 586.4 kg in the control group and 581.5 kg in the treatment group (P value = 0.89). After slaughter, mean warm carcass weight was significantly higher in the treatment groups compared with the controls (treatment = 352.5 kg ± 1.4; control = 347.5 kg ± 1.4; P = 0.01).

[0109] Example 6 Steer The experiment was conducted on a calf farm with 21-day-old crossbred male calves (Holstein crossed with Angus). A total of 33 male calves were randomly assigned as control calves and 42 calves were assigned to receive the novel fatty acid treatment. Calves assigned to the novel fatty acid treatment received a total dose of 10 mL, which was applied topically to the nuchal skin (5 mL) and to the skin above the muzzle (5 mL). The treatment was applied 1 hour before the animals were surgically castrated. Control animals did not receive any treatment and were housed separately to avoid contact with the treatment groups. The novel fatty acid composition (10% fatty acid mixture and 90% mineral oil) was prepared as detailed in Example 1, Table 3. One hour after the calves in the treatment groups were treated with the novel fatty acid composition, all calves were surgically castrated by a licensed veterinarian. To quantify the stress and pain experienced by the study animals, a blood sample was collected 1 hour before castration (baseline sample) and a second blood sample was collected 4 hours after castration. Blood samples were refrigerated immediately after collection and centrifugation. Serum was collected and stored in a -80°C freezer until ELISA assay was performed. Quantification of substance P was measured by a substance P ELISA kit (ADI-900-018A, Enzo Life Science, Inc.). All serum samples were diluted 1:150 with assay buffer immediately before use. Reagent preparation and assay procedures were performed according to the manufacturer's instructions. Cortisol was also measured by an enzyme immunoassay kit; serum samples were diluted 10-fold with the assay diluent provided in the salivary cortisol kit (1-3002, Salimetrics LLC). The procedures followed the manufacturer's instructions.

[0110] Animals receiving the novel fatty acid composition treatment had significantly lower levels of stress associated with the castration procedure, as evidenced by significantly lower levels of cortisol 4 hours after castration (Figure 1). In addition, animals receiving the novel fatty acid composition treatment experienced lower levels of pain, as evidenced by significantly lower levels of substance P 4 hours after castration (Figure 2). Furthermore, treatment with the novel fatty acid mixture enhanced weight gain 17 days after castration (Figure 3).

[0111] Example 7 Dehorned calves The experiment was conducted with 21-day-old Holstein dairy calves in a calf farm. A total of 37 male calves were randomly assigned as control calves and 32 calves were assigned to receive the novel fatty acid treatment. Calves assigned to the novel fatty acid treatment received a total dose of 10 mL, which was applied topically to the nuchal skin (5 mL) and to the skin above the muzzle (5 mL). The treatment was applied 1 hour before the animals were surgically dehorned. Control animals did not receive any treatment and were housed separately to avoid contact with the treatment groups. The novel fatty acid composition (10% fatty acid mixture and 90% mineral oil) was prepared as detailed in (Example 1, Table 3). One hour after the calves in the treatment groups were treated with the novel fatty acid composition, all calves were surgically dehorned by a licensed veterinarian. To quantify the stress and pain experienced by the study animals, a blood sample was collected 1 hour before dehorning (baseline sample) and a second blood sample was collected 4 hours after dehorning. Blood samples were refrigerated immediately after collection and centrifugation. Serum was collected and stored in a -80°C freezer until ELISA assay was performed. Quantification of substance P was measured by a substance P ELISA kit (ADI-900-018A, Enzo Life Science, Inc.). All serum samples were diluted 1:150 with assay buffer immediately before use. Reagent preparation and assay procedures were performed according to the manufacturer's instructions. Cortisol was also measured by an enzyme immunoassay kit. Serum samples were diluted 10-fold with the assay diluent provided in the salivary cortisol kit (1-3002, Salimetrics LLC). The procedure followed the manufacturer's instructions.

[0112] Animals receiving the novel fatty acid composition treatment had significantly lower levels of stress associated with the dehorning procedure, as evidenced by significantly lower levels of cortisol 4 hours after dehorning (Figure 4). In addition, animals receiving the novel fatty acid composition treatment experienced lower levels of pain, as evidenced by significantly lower levels of substance P 4 hours after dehorning (Figure 5). Furthermore, treatment with the novel fatty acid mixture enhanced weight gain 17 days after dehorning (Figure 6).

Claims

1. A fatty acid composition comprising 0.5%-42.8% (wt% / wt%) oleic acid, 0.5%-52.2% (wt% / wt%) myristic acid, 3%-31.2% (wt% / wt%) palmitic acid, 0.5-25 (wt% / wt%) arachidonic acid and derivatives thereof, dissolved in mineral oil.

2. 10. The composition of claim 1 further comprising 0.5%-42.8% (wt% / wt%) oleic acid and at least one of 0.5%-52.2% (wt% / wt%) myristic acid, 3%-31.2% (wt% / wt%) palmitic acid, 0.5-25 (wt% / wt%) arachidonic acid, 0.2%-12.5% ​​(wt% / wt%) lauric acid, 0.1%-22.6% (wt% / wt%) capric acid, 8%-65% (wt% / wt%) linoleic acid, 0.1%-12.6% (wt% / wt%) behenic acid, and derivatives thereof, dissolved in mineral oil.

3. The composition of claim 1 further comprising non-toxic filler compounds (5% to 99%).

4. 4. The composition of claim 3, wherein the non-toxic filler compound is selected from the group consisting of alcohols, amines, squalene, glycerol, caproic acid, azelaic acid, and propionic acid.

5. A composition comprising 65% to 90% of a mineral oil and 10% to 35% of the composition of claim 1.

6. A composition comprising 65% to 99% mineral oil and 1% to 35% of the composition of claim 1.

7. A composition comprising 65% to 90% of a mineral oil and 10% to 35% of the composition of claim 2.

8. A composition comprising 65% to 99% mineral oil and 1% to 35% of the composition of claim 2.

9. The composition of claim 1 in the form of a spray, shampoo, aerosol, or sustained release matrix.

10. The composition of claim 2 in the form of a spray, shampoo, aerosol, or sustained release matrix.

11. 10. The composition of claim 1, which is attached to a chemical carrier molecule selected from the group consisting of a resin, a liposome, a crown compound, and a carrier protein.

12. 10. The composition of claim 1, further comprising 5% to 35% (wt% / wt%) of an enhancer composition comprising amines and fatty acids from indole derivatives, esters of these amines and fatty acids, and ketones selected from acetone, alcohols, and sterols.

13. 10. A method for preventing or treating a condition in a mammal selected from 1) retained placenta, 2) metritis, 3) mastitis, 4) lameness, 5) pain, 6) stress, and 7) diarrhea, comprising administering a pharma- ceutical effective amount of the composition of claim 1 or claim 2 to a mammal in need thereof.

14. The method of claim 13, wherein the composition is in the form of a spray, shampoo, aerosol, or sustained release matrix.

15. 14. The method of claim 13, wherein the composition is applied to the skin of the mammal or to a wall, air, or to a toy in the vicinity of the mammal.

16. A method for treating a condition in a mammal caused by stress, the condition being selected from the group consisting of anxiety, aggressive behavior, weight loss, increased mortality, increased morbidity, and infection, comprising administering a pharma- ceutical effective amount of a composition described in claim 1 or claim 2 to a mammal in need thereof.

17. A method for minimizing weight loss in a live mammal during its transport, comprising administering a pharma- ceutical effective amount of the composition of claim 1 or claim 2 to a mammal in need thereof.

18. 13. A method for increasing warm carcass weight in a mammal during transport to a slaughterhouse, comprising administering a pharma- tically effective amount of the composition of claim 1 or claim 2 to a mammal in need thereof.

19. 13. A method for alleviating pain experienced by a mammal being subjected to a painful procedure, comprising administering to a mammal in need thereof a pharma- ceutical effective amount of the composition of claim 1 or claim 2, wherein the procedure is castration, dehorning, or accidental physical injury.

20. 13. A method for alleviating the stress experienced by a mammal being subjected to a painful procedure, comprising administering to a mammal in need thereof a pharma- ceutical effective amount of the composition of claim 1 or claim 2, wherein the procedure is castration, dehorning, or accidental physical injury.

21. 10. A method for enhancing weight gain in a mammal following the performance of a painful procedure, comprising administering a pharma- tically effective amount of the composition of claim 1 or claim 2 to a mammal in need thereof, wherein the procedure is castration, dehorning, or accidental physical injury.

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