Patent Application
20100034901
Lameness is a major cause of culling and death in female pigs of breeding age, affecting over 20 million animals annually. At least 3 to 10% of young growing swine die or are culled due to lameness. Osteochondrosis (OC) is a major factor in this lameness, causing economic losses potentially exceeding $200 million in the United States alone.
OC is a non-infectious disease of cartilage affecting young growing animals and humans. OC is characterized by abnormal development of articular cartilages of the joints and in the growth plates of the bones, with associated changes in bone development. Lameness occurs when OC changes cause pain and/or interfere with normal skeletal function.
OC is the major cause of lameness in swine. It has been reported that 20 to 80 percent or more of growing pigs are affected by OC. OC severe enough to cause lameness is observed in 5 to 10 percent of horses and large breed dogs, and in 1 of 40 humans. OC is also reported in young growing cattle, especially bulls, and in sheep. OC is not common in cats but has been reported.
In humans, OC primarily afflicts adolescents, an age group that is very physically active and has bones which are still growing. The disease is more common among boys than girls. In children between the ages of 10 to 15, the disease frequently appears at the elbow, knee, or foot joints. Afflicted humans experience tenderness, swelling, and pain at the affected joints which worsens with activity.
Among the more common forms of OC in human children are: Freiberg's disease, which occurs in the head of the metatarsals of the feet in children between the ages of 12-15; Legg-Calve-Perthes' disease, which occurs in the hip in children between the ages of 6 to 9; Osgood-Schlatter disease, which occurs in the tibial tubercle apophysis at the insertion of the patellar tendon in the knee in children between the ages of 10 to 15; Panner's disease, which occurs in the capitellum of the distal humerus at the elbow in children between the ages of 5-10; and Sinding-Larsen-Johannson disease, which occurs at the inferior pole of the patella in the knee in children between the of ages 10-15. Animal correlates of each human OC condition are observed, with species and breed related “predilection sites”. In particular, different specific joints are more likely to be affected in a given species or breed; for instance there is a tendency for “elbow dysplasia” to develop in German Shepherds.
Thus, improved methods for the prevention and treatment of OC would be of great economic value in the livestock industry, would reduce animal suffering, and would help alleviate the painful joint discomfort and loss of function and mobility experienced by humans suffering from this disease.
Furthermore, phosphate pollution resulting from excess phosphorus in animal feed is an increasing problem. Such phosphorus can potentially contaminate ground water. There is a need to provide animal feed with reduced phosphorus content to reduce ground water contamination. Reduction in phosphorus use would assist animal producers in complying with nutrient control regulations.
Leg weakness problems in commercial turkey production is a major cause of economic loss. Above average commercial producers may average 12-13% mortality with 5.5-6% of that mortality coming in the last 5 weeks of production. This late mortality is very expensive to the industry since much of the expense of raising the bird has occurred by that time. The top 24 turkey growers in the US lost between 23 and 32 million turkey's per year to mortality losses and leg weakness problems are a significant cause of those losses. The cost of this loss is estimated at $131 million dollars/year or above.
United States Department of Agriculture (USDA) figures updated 29 Nov. 2007 indicate turkey production for 2007 is estimated at 262 million birds with an average live weight at market of 28.31 pounds for a total production of 7.4 billion pounds with a market value of USD $4.0 billion. That is up $0.5 billion from 2006 due to the increased per pound market value which is $0.54 per pound for 2007. The average value of a market weight turkey is estimated at $15.28 for 2007 (interestingly the total value of turkey production in the USA is greater than the combined values of rice, peanuts, and tobacco production).
A dataset of 88,701 tom turkeys raised from 1 day of age to market at 17 to 19 weeks showed an overall total mortality of 12,379 birds or 14.0%. This data is typical or better for mortality rates for the national commercial turkey industry, for which the national average mortality is not known exactly but sometimes said to be 15 to 16%. Approximately half of the mortality (6191 birds or 6.967%) occurred after the birds reached 12 weeks of age, with 4543 birds or 5.1% dying after 14 weeks of age which is only 3 to 5 weeks before marketing. Almost all of the post-12 week mortality can be attributed to the leg weakness syndrome.
A total mortality of 14% is equivalently expressed as an 86% survivability rate. To achieve a market production of 262 million birds, 304.6 million day 1 poults must be placed. The expected mortality after week 12 is 7% or 21.3 million birds. Since those birds at 12 weeks of age are ⅔ of the market age of 18 weeks, it is reasonable to assign a value of $10 per head for a mortality cost of $213 million nationally. Further, production losses are not limited to mortality. Significant additional economic loss occurs due to lameness and poor performance (morbidity) among affected birds.
Irregular metabolism in the growth plate area in poultry is associated with dwarfism and tibial dyschondroplasia (TD). Tibia dyschondroplasia is a disease associated with rapid growth rate, and genetic selection for growth has actually resulted in the increased incidence of this skeletal disease. Traditional approaches have been unable to reduce the occurrence of abnormal bone growth substantially.
In addition, other factors that lead to lameness and leg weakness are: (1) lesions in the cartilage and subarticular bone; (2) necrosis of the joint surface and subarticular bone, which can measured by the number of infarcts (necrotic tissue caused by the obstruction of the local blood supply); (3) growth plate widening, which can lead to hyperplasia, necrosis and hemorrhage; and (4) articular cartilage damage, which can be evidenced by reduced concentrations of glycosaminoglycans (GAGs), hydroxyproline, and other biomolecules that are related to or components of proteoglycans or collagen.
It would be highly advantageous if one could provide a reduced phosphorus animal feed that would simultaneously facilitate the prevention and treatment of OC, TD, Leg weakness, and lameness.
The inventors have made the unexpected discovery that the administration of boron containing compounds and vitamin C is effective in preventing and treating leg weakness and lameness in animals. In one embodiment, this invention provides an animal feed containing supplemental boron and vitamin C. Animal feeds contain plant material. Boron is a required element for plant growth. As such all plants and hence all plant material in animal feeds contain some boron, e.g. 10-20 ppm boron in corn/soybean feed (unless the boron as been extracted). In addition, many plants contain vitamin C. The animal feeds of the present invention contain supplemental boron in addition to the boron naturally present in the animal feed from the plant material and supplemental vitamin C in addition to the vitamin C naturally present in the animal feed from the plant material. The supplemental boron is supplied as a boron-containing compound, as plant material with elevated boron levels or as microorganisms such as yeast with elevated boron levels. Among the boron-containing compounds that may be used in the practice of the present invention are sodium borate and boric acid as typical boron sources. However, the invention is not limited to these forms of boron. Also included are other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen containing borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate.
Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C.
In this embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 500 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 1000 ppm. In other embodiments, the boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 150 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 500 ppm. In yet another embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 50 ppm or about 25 to 50 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 25 to about 500 ppm. Among the animals that would benefit from the animal feed are humans, birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, and cats.
In a further unexpected discovery, the inventors have determined that the addition of supplemental boron to animal feed allows for the reduction in phosphorus content of the animal feed. Thus, in another embodiment, the invention provides an improved animal feed containing supplemental boron containing compounds, vitamin C, and reduced phosphorus content. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid. However, the invention is not limited to these forms of supplemental boron. Other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used as well. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen containing borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among these organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. The boron can be combined with talc in a ratio of boron containing compound to talc of approximately 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1; 21:1; 22:1, 23:1; 24:1 or 25:1 prior to addition to the animal feed. The supplemental boron containing compounds are included in the animal feed at about 1 to about 500, about 1 to about 150 or about 50 ppm or about 25 to 50 ppm supplemental boron and the total phosphorus content is reduced by at least 3% as compared to a comparable animal feed without supplemental boron. Generally, the animal feed is supplemented with boron at concentrations ranging from about 5 to about 150 ppm. The animal feed is suitable for humans, birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, and cats among other animals.
In another embodiment, the invention provides a method of decreasing the amount of phosphorus excreted by an animal. In this embodiment, animals are fed an animal feed with an improved animal feed composition containing about 1 to about 500, about 1 to about 150 or about 50 ppm or about 25 to 50 ppm supplemental boron supplied as boron containing compounds, plant material with elevated boron levels, yeast or other microorganisms with elevated boron levels, and about 1 to about 1000, about 1 to about 500 or about 50 ppm or about 25 to 50 ppm supplemental vitamin C, in which the animal feed composition has at least a 3% reduction in phosphorus as compared to a comparable animal feed without supplemental boron and vitamin C. Generally, the animal feed contains supplemental boron at concentrations ranging from 5-150 ppm. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid can be used. However, other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. The method is suitable for use with humans, birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, and cats among other animals.
An additional embodiment provides a method of increasing the efficiency of absorption of phosphorus in animals. In this embodiment, animals are fed an animal feed with an improved animal feed composition containing about 1 to about 500, about 1 to about 150 or about 50 ppm or about 25 to 50 ppm supplemental boron supplied as boron containing compounds, plant material with elevated boron levels, yeast or other microorganisms with elevated boron levels, and about 1 to about 1000, about 1 to about 500 or about 50 ppm or about 25 to 50 ppm supplemental vitamin C, in which the animal feed composition has at least a 3% reduction in phosphorus as compared to a comparable animal feed without supplemental boron and vitamin C. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid. However, other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen containing borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. The method is suitable for use with humans, birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, and cats among other animals.
In yet an additional embodiment, this invention provides a method of reducing environmental phosphorus pollution from an animal farm. In this embodiment, animals are fed an animal feed with an improved animal feed composition containing about 1 to about 500, about 1 to about 150 or about 50 ppm or about 25 to 50 ppm supplemental boron supplied as boron containing compounds, plant material with elevated boron levels, yeast or other microorganisms with elevated boron levels, and about 1 to about 1000, about 1 to about 500 or about 50 ppm or about 25 to 50 ppm supplemental vitamin C, in which the animal feed composition has at least a 3% reduction in phosphorus as compared to a comparable animal feed without supplemental boron and vitamin C. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid. However, inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen containing borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. The method is suitable for use with humans, birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, and cats among other animals.
In a further embodiment, the invention also provides a method of treating or preventing OC by administering a therapeutically effective amount of a boron containing compound and vitamin C to an animal in need of such treatment. In such an embodiment, the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. The boron containing compound can be sodium borate or boric acid. However, the invention can be used with other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen containing borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. The supplemental boron and vitamin C containing compounds can be administered prior to the appearance of symptoms of osteochondrosis as a preventive measure. Among the animals that can benefit from this invention are humans, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, cats, and birds.
In a further unexpected discovery, the inventors have determined that the addition of supplemental boron and vitamin C to animal feed allows for the reduction in the incidence and extent of lesions in the cartilage and subarticular bone of animals. Thus, in this embodiment, the invention provides a method for reducing the incidence and extent of lesions in the cartilage and subarticular bone of animals. In this embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 500 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 1000 ppm. In other embodiments, the boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 150 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 500 ppm. In yet another embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 50 ppm or about 25 to 50 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 25 to about 500 ppm. In such an embodiment, the feed could also be in the form of a liquid. In such an embodiment, the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid can be used. However, other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the animals that would benefit from the animal feed are birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, cats, as well as humans.
In a further unexpected discovery, the inventors have determined that the addition of supplemental boron and vitamin C to animal feed allows for the prevention of necrosis of the joint surface and subarticular bone as measured by the number of infarcts (necrotic tissue caused by the obstruction of the local blood supply). Thus, in this embodiment, the invention provides a method for preventing necrosis of the joint surface and subarticular bone as measured by the number of infarcts (necrotic tissue caused by the obstruction of the local blood supply). In this embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 500 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 1000 ppm. In other embodiments, the boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 150 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 500 ppm. In yet another embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 50 ppm or about 25 to 50 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 25 to about 500 ppm. In such an embodiment, the feed could also be in the form of a liquid. In such an embodiment, the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid can be used. However, other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the animals that would benefit from the animal feed are birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, cats, as well as humans.
In a further unexpected discovery, the inventors have determined that the addition of supplemental boron and vitamin C to animal feed allows for the reduction of hyperplasia, necrosis and hemorrhage. Thus, in this embodiment, the invention provides a method for reducing hyperplasia, necrosis and hemorrhage. In this embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 500 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 1000 ppm. In other embodiments, the boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 150 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 500 ppm. In yet another embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 50 ppm or about 25 to 50 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 25 to about 500 ppm. In such an embodiment, the feed could also be in the form of a liquid. In such an embodiment, the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid can be used. However, other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the animals that would benefit from the animal feed are birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, cats, as well as humans.
In a further unexpected discovery, the inventors have determined that the addition of supplemental boron and vitamin C to animal feed allows for the reduction of dysplasia (abnormal development and/or abnormal structure) of cartilage, growth plate and bone as measured by the growth plate width (a wide growth plate being representative of abnormal growth and improper ossification). Thus, in this embodiment, the invention provides a method for reducing dysplasia (abnormal development and/or abnormal structure) of cartilage, growth plate and bone as measured by the growth plate width; (a wide growth plate being representative of abnormal growth and improper ossification). In this embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 500 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 1000 ppm. In other embodiments, the boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 150 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 500 ppm. In yet another embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 50 ppm or about 25 to 50 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 25 to about 500 ppm. In such an embodiment, the feed could also be in the form of a liquid. In such an embodiment, the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid can be used. However, other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the animals that would benefit from the animal feed are birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, cats, as well as humans.
In a further unexpected discovery, the inventors have determined that the addition of supplemental boron and vitamin C to animal feed allows for the reduction articular cartilage damage as evidenced by higher concentrations, typical of healthy tissue, of glycosaminoglycans (GAGs), hydroxyproline, and other biomolecules that are related to or components of proteoglycans or collagen. Thus, in this embodiment, the invention provides a method of reducing articular cartilage damage as evidenced by higher concentrations, typical of healthy tissue, of glycosaminoglycans (GAGs), hydroxyproline, and other biomolecules that are related to or components of proteoglycans or collagen. In this embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 500 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 1000 ppm. In other embodiments, the boron containing compounds are typically included in animal feed at concentrations providing about 1 to about 150 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 1 to about 500 ppm. In yet another embodiment, the supplemental boron containing compounds are typically included in animal feed at concentrations providing about 50 ppm or about 25 to 50 ppm supplemental elemental boron and the vitamin C containing compounds are at concentrations providing about 25 to about 500 ppm. In such an embodiment, the feed could also be in the form of a liquid. In such an embodiment, the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid can be used. However, other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the animals that would benefit from the animal feed are birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, cats, as well as humans.
In another embodiment, the invention provides a method of decreasing the amount of pre-weaning mortality in animals. In another embodiment, the invention provides a method of improving reproductive rates of animals by increasing the rate of return to estrus and conception rates. In these embodiments, previously pregnant, pregnant, nursing and/or lactating animals are fed a diet of increased boron. The diet may contain about 1 to about 500, about 1 to about 150 or about 50 ppm or about 25 to 50 ppm supplemental boron containing compounds and about 1 to about 1000, about 1 to about 500 or about 50 ppm or about 25 to 50 ppm supplemental vitamin C. The boron and vitamin C may be provided in improved animal feed composition or in milk or water. Generally, the milk, water or animal feed contains supplemental boron and vitamin C at concentrations ranging from 5-150 ppm. In such embodiments, the supplemental boron containing compound can be sodium borate or boric acid can be used. However, other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among the organic forms are complexes and compound formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C. The method is suitable for use with humans, birds, pigs, horses, mules, donkeys, cattle, sheep, goats, llamas, dogs, and cats among other animals.
In an additional embodiment, the boron and vitamin C containing compounds are added to drinking water, mineral or vitamin supplements, in a milk formulation, or other food products for the treatment and prevention of TD, leg weakness, OC and/or reduction in pre-weaning mortality.
In an additional embodiment, this invention provides a vitamin C, boron, and talc composition where the ratio of boron-containing compound to talc is approximately 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1; 21:1, 22:1, 23:1, 24:1 or 25:1. In such an embodiment, the boron is a boron containing compound which can be sodium borate or boric acid. However, the invention is not limited to these forms of supplemental boron. Other inorganic forms of boron such as calcium borate, as well as, organic boron compounds and complexes that dissociate or are metabolized in the body to release boron as borate or boric acid can be used as well. Among the inorganic forms are sodium borate, boric acid, calcium borate, magnesium borate, halogen containing borate, ammonium borate, potassium borate, iron and magnesium containing borate, tantalum borate, beryllium borate, iron and nickel containing borate, carbonate containing borate, sodium and calcium containing borate, arsenate containing borate, calcium and rare earth containing borate, sulphate containing borate, magnesium and calcium containing borate, manganese borate, aluminum borate, calcium and strontium containing borate, phosphate containing borate, tin borate, strontium borate, zinc borate, calcium borosilicate, sodium borosilicate, aluminum borosilicate, calcium and rare earth containing borosilicate, lead borosilicate, barium borosilicate, lithium borosilicate, and sodium fluoroborate. Among these organic forms are complexes and compounds formed by boron, usually as boric acid, with fructose, sorbitol, mannitol, xylitol, sorbose, threonine, methionine, modified starches, hydrolyzed starches, oxidized starches, non-modified starches, dextrins, amidated sugars, glucosamine, mannosamine, esters of glycerol fatty acids, salicylate complexes, salts of bisoxalato acid, calcium borosucrose, alcohols, alcohol amines, sugar acids, saccharic acid, gluconic acid, aminated sugar acids, and calcium borogluconate. Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C.
Talc is available for use in the present exemplary embodiments from a variety of commercial sources. For example, Luzenac America is a supplier of talc. Examples of talc products from Luzenac America include: E-Z Flow 40, E-Z-Flow MB, E-Z Flow MT, E-Z Flow RM, and E-Z Flow VT.
Boron has long been known to be an essential plant nutrient, but a role for boron in human physiology has only recently been appreciated. Furthermore, vitamin C is well known to have beneficial effects on animal physiology. The present inventors have discovered a beneficial effect of boron and vitamin C supplementation of animal and human diets. In particular, although previous studies had shown that boron containing compounds could alleviate the bone disease, osteoporosis, the present inventors have discovered that boron and vitamin C containing compounds also alleviate a disease of the joints and growth plate cartilages, osteochondrosis (OC), and Tibial dyschondroplasia (TD).
Osteoporosis is a disease in which bones become fragile and become increasingly likely to break as the disease progresses. Osteoporosis, or porous bone, is characterized by low bone mass and structural deterioration of bone tissue, which leads to bone fragility and an increased susceptibility to fractures of the hip, spine, and wrist. Thus, osteoporosis is a disease that specifically strikes the bone, generally after full and normal development. Also, because of its progressive nature, osteoporosis is a disease that most commonly manifests itself in older individuals. One out of every two women and one in four men over the age of 50 will have an osteoporosis-related fracture in their lifetimes.
In contrast, osteochondrosis is a generalized skeletal disease of growing animals and results from a disturbance in the articular and growth plate cartilages. The bone is only secondarily affected. As a consequence, dyschondroplasia is technically a more correct term to describe this condition. A further condition, osteochondrosis dissecans, results in the chipping, fracturing and/or fragmentation of the articular surface. Osteochondrosis dissecans is thought to be caused by an underlying weakness in the cartilage caused by an osteochondrotic lesion. Lesions are characterized by focal impaired endochondondral ossification, resulting in areas of retained cartilage extending into the subchondral bone. See R. John Wardale and Victor C. Duance. Journal of Cell Science 107, 47-59 (1994).
Because of the differences in the pathobiology of OC and osteoporosis—the two diseases affect different aspects of the bone system and different age groups—one would not expect that a treatment that alleviates osteoporosis, a degenerative disease of the bones of the elderly, would help in the treatment of OC, a disease of the cartilage in joints of the young. The inventors have found that surprisingly, boron containing compounds are useful agents in the prevention and treatment of OC.
Tibia dyschondroplasia is a disease associated with rapid growth rate, and genetic selection for growth has actually resulted in the increased incidence of this skeletal disease. Traditional approaches have been unable to reduce the occurrence of abnormal bone growth substantially. Irregular metabolism in the growth plate area in poultry is associated with dwarfism and tibial dyschondroplasia (TD).
The extracellular matrix (ECM) of the articular cartilage provides cushioning between opposing bone surfaces at a joint in a mammalian limb. Synovial fluid is the fluid contained in joints. Synovial membranes line the joints, bursae, and tendon sheaths. The function of the synovial fluid is to lubricate the joint space and transport nutrients to the articular cartilage. The articular cartilage provides a low friction point of contact for the smooth flexing operation of the joints and also a cushioning function at joints, by absorbing the impact of shocks transmitted through the bones and supporting the weight of the animal. The cartilage is composed of a variety of components including proteoglycan and a collagen network in an aqueous environment.
Proteoglycans play a role in maintaining the cushioning seen at joints. The cartilage ECM is illustrated as a network of the collagen fibers which interlocks with and is interlocked by proteoglycan. The proteoglycan is a flexible gel-like material and the collagen forms a mesh-network that holds the proteoglycan in place. The proteoglycan provides compressive strength while tensile strength is provided by the collagen network. Proteoglycan in articular and growth plate cartilage contains large amounts of sulfated glycosaminoglycans (GAG) that have a strong negative charge. At physiologic pH, these negatively charged GAG molecules draw sodium ions and water into the ECM of the cartilage, causing the proteoglycan to “inflate”. The inflated proteoglycan provides buoyant pressure to resist compression, thus protecting the collagen network and underlying structures from compression damage. Better cushioning and thus greater compression resistance is provided by a fully hydrated proteoglycan complement and a fully extended and taut collagen network.
Without limiting this invention to any particular mechanism nor being bound by theory, one potential model for how boron functions in OC is the “standing hypothesis”. In this hypothesis, boron functions by crosslinking the proteoglycan into the extracellular matrix. One postulated mechanism for how this occurs is that boron provides for three-dimensional boroester crosslinking of carbohydrate, proteoglycan, glycoprotein, glycolipid, lipid, protein, and amino acid structures. In the case of extracellular membrane structures such as cartilage and neural tissue, this would include proteoglycans such as aggrecan (the large aggregating proteoglycan of cartilage), complex proteins such as collagen in its various forms and types, and associated proteins such as cartilage link protein. The crosslinking of the proteoglycan stabilizes and unifies the matrix, allowing for better distribution of compressive forces and prevention of proteoglycan loss, which would decrease the cushioning ability of the synovial membrane. In contrast, boron functions to prevent osteoporosis by increasing the plasma levels of hydroxylated steroids. See U.S. Pat. No. 4,849,220. Thus, one would not have predicted that boron would have an effect in treating a disease of the cartilage, such as OC, which has a totally different etiology from osteoporosis. In osteoporosis, the bone itself is directly affected. In OC, the cartilage is affected.
Although the precise cause of OC is not yet known, a number of mechanisms for the progression of this disease have been suggested. The influence of compressive forces in producing damage to the growing and transitioning cartilage appears to be a major factor. Studies in pigs have suggested that focal changes in blood supply during normal epiphyseal growth is central to the pathogenesis of osteochondrosis. Cartilage canals are temporary blood vessel-containing structures within growing cartilage. The canals gradually regress with age during the process of chondrification, wherein the blood vessels contained within the canals are replaced with cartilage. Formation of the lesions associated with osteochondrosis has been associated with the premature chondrification and regression of these canals. In particular, the premature disruption of the blood supply results in necrosis of the cartilage canal distal to the point of interruption. See Ytrehus et al. Bone 35: 1294-1306 (2004). Thus, it is not surprising that severe clinical osteochondrosis appears most commonly in fast-growing animals with rapid weight gain. See Wardale and Duance Journal of Cell Science 107: 47-59 (1994).
It has also been demonstrated in humans and dogs that in osteochondrosis, the proteoglycan of cartilage is resorbed by the action of matrix metalloproteinase-3 (MMP-3) derived from synovial membrane cells and chondrocytes. See Shinmei et al. 1991; Okada et al. 1992; Mehraban et al. 1994. Loss of proteoglycan from the extracellular matrix of the cartilage would lead to a decreased capacity of the cartilage to absorb and cushion compressive forces.
In order to further probe the contribution of boron deficiency in OC, and to test whether boron functions in OC through quadrivalent crosslinking, 3-nitrophenylboronic acid (3-NPB) was administered to animals. 3-NPB blocks crosslinks by binding to sites normally occupied by boric acid or borate. The results are described in Example 5. 3-NPB treated animals had increased lameness and clinical manifestations of OC. The increase in lameness could be prevented by supplementing the diet with boron. These experiments show that OC is directly correlated to boron levels in pigs, horses, cattle, and dogs.
Given the widespread occurrence of OC in livestock and in particular, pigs, this invention discloses a safe and effective means of preventing and treating OC by providing for animal feed to be supplemented with boron and vitamin C containing compounds. It will be appreciated by one of skill in the art that animal feeds, derived at least in part from plant materials, will contain basal levels of boron. For instance, typical alfalfa contains about 37 ppm boron. Diets fed to livestock are generally quite low in vitamin C, however, a diet based on leafy green plants will provide some vitamin C. Furthermore, most domestic animals have significant ability to synthesize vitamin C metabolically from glucose. The ability of that synthetic process to meet demand for vitamin C is dependent upon the demand for growth, reproduction, antioxidant status, and the synthesis capacity of the animal under varying conditions. Thus, the term supplemental boron and vitamin C as used herein refers to exogenously added boron and vitamin C that supplements the basal levels of boron and vitamin C already present in the animal's diet and metabolism. When the term boron is used in this disclosure, it can denote both elemental boron and boron containing compounds. The boron containing compounds useful for the practice of this invention may include any suitable organic or inorganic boron containing compounds, including boron containing minerals. Among the preferred forms of boron are sodium borate and boric acid. Other useful inorganic forms of boron include calcium borate. One of skill in the art will recognize that other inorganic forms of boron that may be used in this invention include borates with: magnesium, halogen, ammonium, potassium, iron and magnesium, tantalum, beryllium and nickel, carbonate, sodium and calcium, arsenate, calcium and rare earth, sulphate, magnesium and calcium, manganese, aluminum, calcium and strontium, phosphate, tin, zinc, and strontium. Other forms include: borosilicates or silicoborates with calcium, sodium, aluminum, calcium and rare earth, lead, barium, lithium, and fluoroborate with sodium. Natural inorganic boron containing compounds are known to skilled artisans by various mineral names such as borax, colemanite, hydroboracite, kemite, ulexite, datolite, danburite, szaibelyite, suanite, inderite, sassolite, inyoite, probertite, howlite, ezcurrite, kurnakovite, meyerhofferite, priceite, nobleite, and searlesite to name but a few such designations. A listing of inorganic borate compounds and minerals can be found in Supplement to Mellor's Comprehensive Treatise on Inorganic and Theoretical Chemistry, Volume V Boron, by Joseph William Mellor, Longman Group Limited, London, 1980.
When the term vitamin C is used in this disclosure, it can denote vitamin C as any mix of ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate. However, the invention is not limited to these forms of vitamin C.
Examples of organic boron-containing compounds are well known to those of skill in the art. Examples of such organic boron-containing compounds are found in U.S. Pat. Nos. 4,312,989, 4,499,082, and 5,312,816 all of which are hereby incorporated by reference. Among the forms of organic boron that would be useful in the practice of this invention are organic boron complexes such as boron threonine, boron methionine, and boron ascorbate, as well as boron complexed with other amino acids. These amino acids can include the 20 common amino acids that are specified by the genetic code, as well as variant and modified amino acids which are not encoded by the genetic code. These are examples of organic forms of boron that are rapidly metabolized to release borate or boric acid. Other useful forms of organic boron are boron carbohydrate complexes such as those disclosed in U.S. Pat. No. 5,962,049. Among the carbohydrates that form useful complexes with boron include saccharides such as fructose, sorbitol, mannitol, xylitol, and sorbose. A commercially available form of boron complexed with fructose is Fruitex B™ available from FutureCeuticals and described in U.S. Pat. No. 5,962,049.
Other organic forms of boron that can be used in the practice of this invention include: borated modified starches (such as hydrolyzed or oxidized starches), borate non-modified starches, borated dextrins, borated amidated sugars (such as glucosamine or mannosamine), borate esters of glycerol fatty acids, borate-salicylate complexes, salts of bisoxalato borate (such as sodium or potassium salts), calcium borosucrose, borate esters (such as (RO) 3B), alcohol amine borate esters, and borate complexes with sugar acids (such as saccharic acid and gluconic acid), and borate complexes with aminated sugar acids. One particularly desirable sugar acid to use in this invention is calcium borogluconate. Yet another form of boron are anion exchange resins which can be boronated. One such resin which can be boronated is Amberlite™.
It will be appreciated by one of skill in the art that when a particular boron and vitamin C containing compound is described herein, it is intended that all possible solvates, pharmaceutically acceptable salts, esters, amides, complexes, chelates, stereoisomers, geometric isomers, crystalline or amorphous forms, metabolites, metabolic precursors or prodrugs of the compound are also separately described by a chemical structural formula or chemical name. Furthermore, if any of the boron and vitamin C containing compounds described herein contain stereochemistry, all enantiomeric and diastereomeric forms of the compound are intended. Thus, when applicable, boron and vitamin C containing compounds may occur as racemates, racemic mixtures and as individual diastereomers, or enantiomers with all isomeric forms being included. A racemate or racemic mixture does not necessarily imply a 50:50 mixture of stereoisomers.
Furthermore, it will be appreciated by one of skill in the art that the borates and ascorbates of the present invention will encompass many different grades, including those that are FDA and non-FDA approved. Thus, among the grades of borates and ascorbates that can be used in the practice of this invention are: pharmaceutical or formulary grade, nuclear grade, fertilizer grade, industrial grade, pesticidal grade, and special quality (SQ) grade.
Suitable ranges for use of the boron and vitamin C containing compounds includes the supplementation of boron and vitamin C in animal feed from about 1 to about 500 ppm for boron and about 1 to about 100 ppm for vitamin C, above that naturally present in the animal feed. Another suitable range for supplementation is about 1 to about 150 ppm for boron and about 1 to about 500 ppm for vitamin C. As shown in
The boron described herein may be combined with talc. The boron-containing compound to talc ratio may be approximately 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1; 21:1, 22:1, 23:1, 24:1 or 25:1.
Incorporation of Supplemental Boron and Vitamin C into Animal Feeds
A variety of methods are known in the art for the production of animal feeds. These various methods can be adapted to allow inclusion of supplemental boron and vitamin C into the feed in amounts that will have a beneficial effect on OC and TD when fed to animals.
For example, supplemental boron and vitamin C in the amounts disclosed above can be incorporated into animal feed compositions such as those described in U.S. Pat. No. 3,946,109. Alternatively, a variety of other feed compositions are commercially available from suppliers such as Purina, ADM, Land O'Lakes, and Moorman's. Supplemental boron and vitamin C can be mixed into a composition of choice using for instance, the mixing methods disclosed in U.S. Pat. No. 4,189,240. The composition containing supplemental boron and vitamin C can be used to form animal feed food blocks such as those disclosed in U.S. Pat. No. 5,120,565. Alternatively, supplemental boron and vitamin C can be incorporated into an animal feed composition which is formed by methods such as spray drying as disclosed in U.S. Pat. No. 4,777,240. The citation of these patents is solely to illustrate various methods available in the art for incorporating supplemental boron into an animal feed product and is not meant to limit the practice of the invention to the use of any one or more of these methods.
Other boron sources that can be incorporated into animal feeds to practice this invention include yeast preparations that are high in boron. It is already common practice to incorporate yeast into animal feeds. Hence, it would be fairly straightforward to include yeast with elevated boron levels in animal feed. Alternatively, crops that have been grown in soils with elevated boron levels can be harvested specifically for the purpose of serving as an enhanced boron source that can be incorporated into animal feeds. Such elevated boron levels may be naturally in the soil, may result from boron pollution, or may be added to the soil by fertilization or other means. Alternatively, supplemental boron containing compounds can be added to supplements, base mixes, and premixes that also contain vitamins and minerals. Such supplements, mixes, or premixes are typically added at an amount to constitute 0.5% to 30% of the final animal feed composition. In such an embodiment, the elemental boron concentration would be much higher (from about 3 times to 200 times higher) prior to dilution in the animal feed to result in a supplemental boron equivalent of 1-500 ppm over a total daily ration.
Another alternative is to supplement animal feeds with foods, such as alfalfa, grapes, or coffee grounds, which are naturally high in boron content. Additionally, these and other foods can be manipulated to contain higher levels of boron by growth under elevated boron conditions as described above or by means of transgenic plant technology or other recombinant methods.
In a further embodiment, supplemental boron and vitamin C containing compounds can be provided as dietary supplements that can be directly hand-fed or “top-dressed” onto an animal feed. Such an embodiment could be in a formulation that contains other nutrients, excipients, or flavors. As an example, an equine nutrient supplement containing supplemental boron and vitamin C and other vitamins and minerals could be fed to a horse with a small spoon or cup or in the form of a bar or pellet. Alternatively, the supplement could be placed on top of or mixed in the animal's feed.
Such boron can be supplied to animal feeds as a boron-talc composition. The ratio of boron-containing compound to talc in the boron-talc composition can be approximately 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1; 21:1, 22:1, 23:1, 24:1 or 25:1
Should it be necessary, boron and vitamin C levels can be precisely determined by a variety of methods known in the art. U.S. Patent Publication 20040020840 and the patents disclosed therein describe a number of such methods.
Phosphate pollution resulting from excess phosphorus in animal feed is an increasing problem. For example, approximately 70% of the phosphorus in a typical corn/soybean meal diet is unavailable to pigs, according to the National Research Council's 1998 Nutrient Requirements for Swine. This unavailable phosphorus ends up being excreted in manure. The high phosphate content of swine manure contributes to the environmental pollution associated with pig farming. Reducing the amount of excreted nutrients, particularly phosphorus, in swine production systems is an environmental priority and an important economic issue facing the swine industry. Thus, a means to increase the bioavailability of phosphorus in feed ingredients used to formulate swine rations would be desirable. The inventors have found that inclusion of supplemental boron in pig feed results in an increased absorption and utilization of the phosphorus present in pig diets. Supplemental boron promotes the efficient incorporation of phosphate into the calcium phosphate (hydroxyapatite) of bones. This effect is expected to be true in other animals as well.
By increasing the efficiency of absorption and utilization of phosphorus from animal diets such as pig diets, the inventors have found that the amount of phosphorus in typical pig feed formulations can be reduced. These results are shown in Example 4. The increased utilization of phosphorus from pig diets coupled with the reduction in the starting amount of phosphorus in pig feed can be expected to contribute to a reduction of phosphate pollution that results from pig farming. Thus, the inclusion of supplemental boron in animal feed as taught by this invention will not only contribute to the prevention and treatment of OC, it will also contribute to pollution reduction. While the foregoing discussion has focused on pigs, this invention is not limited to the reduction of phosphorus from pig feed exclusively. Rather, one of skill in the art will recognize that reduction of phosphorus use is applicable to all animals.
The formation of most bones of the axial skeleton begins with the formation of a cartilage model which is calcified and remodeled into bone that is mineralized with calcium and phosphate. Supplemental boron enhances the efficiency of this process. The postulated mechanism by which supplemental boron enhances the efficiency of the process is through stabilization of the extracellular matrix, although there may be other mechanisms.
Supplemental boron improves the efficiency of cartilage transformation/bone mineralization which improves the structural integrity of bone and bone mineralization characteristics. Calcium is added to diets at a level that promotes sufficient bone strength. The level of calcium that promotes optimum bone strength also paradoxically inhibits the intestinal absorption of phosphorus. Phosphorus absorption is also more efficient when dietary phosphorus level is reduced. The inventors have discovered that addition of supplemental boron to animal feed promotes bone mineralization and permits a proportional 3 to 5% reduction of both calcium and phosphorus in the animal feed while maintaining bone strength.
The 1998 NRC report on “Nutrient Requirements of Swine”, data from which is shown below in Table 1, indicates that typical requirements for calcium and phosphorus will vary during the lifetime of a pig. At earlier stages, when the bones of the skeleton are still undergoing development, greater amounts of calcium and phosphorus are needed to support increased bone growth. The requirements for calcium and phosphorus decrease as a pig matures and bone development is completed. Although the data presented below are for pigs, similar trends in requirements for calcium and phosphorus are observed during the life cycle of other animals.
The inclusion of supplemental boron and vitamin C in the diet of various animals allows the levels of calcium and phosphorus to be reduced by at least 3% throughout the life cycle of animals. Thus, while the ratio of calcium to phosphorus is generally kept constant at each weight range indicated in Table 1, the absolute amounts of calcium and phosphorus can be lowered by at least 3% due to the addition of supplemental boron containing compounds.
Thus, in another embodiment, this invention provides an animal feed containing supplemental boron and vitamin C with a reduced level of phosphorus. In one embodiment, the supplemental boron is preferably provided at a concentration of about 1 to about 500 ppm elemental boron and vitamin C at a concentration of about 1 to about 1000 ppm and the phosphorus level is reduced by 3 to 5% as compared to comparable animal feed without supplemental boron. The calcium level is generally reduced comparably to the phosphorus level. However, it will be recognized that if the supplemental boron containing compound is supplied as a calcium salt, such as calcium borate or calcium borogluconate, levels of calcium in the animal feed can also be correspondingly reduced. One such compound, calcium borogluconate, is already in use to treat hypocalcemia in cattle, sheep, and goats. In another embodiment, the supplemental boron concentration is preferably about 1 about 150 ppm, the supplemental vitamin C concentration is preferably about 1 about 500 ppm, and the phosphorus level is reduced by 3 to 5% as compared to comparable animal feed without supplemental boron. In yet another embodiment, the supplemental boron concentration is preferably about 25 ppm to 50 ppm, the supplemental vitamin C concentration is preferably about 25 ppm to 50 ppm, and the phosphorus level is reduced by 3 to 5% as compared to comparable animal feed without supplemental boron.
In another embodiment, the invention provides a method of decreasing the amount of pre-weaning mortality by animals. In this embodiment, pregnant, nursing or lactating animals are fed a diet of an improved animal feed composition containing about 1 to about 500, about 1 to about 150 or about 50 ppm or about 25 to 50 ppm supplemental boron containing compounds and about 1 to about 1000, about 1 to about 500 or about 50 ppm or about 25 to 50 ppm supplemental vitamin C containing compounds in which the animal feed composition has at least a 3% reduction in phosphorus as compared to a comparable animal feed without supplemental boron. Generally, the animal feed contains supplemental boron and vitamin C at concentrations ranging from 5-150 ppm for boron and 25-1000 ppm for vitamin C. In such an embodiment, the supplemental boron containing compound can be sodium borate or boric acid can be used or other inorganic forms of boron as described herein. Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate.
Osteochondrosis with its various manifestations has been found to be strikingly similar in six species of animals in which it has been reported. This has prompted experts to assert that it would be expected that osteochondrosis in humans would have the same etiology, pathogenesis, and pathology as has been observed in animals. See Olsson, S. E. and Reiland, S. 1978. The nature of osteochondrosis in animals—summary and conclusions with comparative aspects on osteochondrosis dissecans in man. Acta Radiologica Supplement No. 358:299-306.
Osteochondrosis in humans is defined in Dorland's Medical Dictionary as follows: a disease of the growth or ossification centers in children that begins as a degeneration or necrosis followed by regeneration or recalcification. Also called epiphyseal ischemic necrosis (q.v.), it may affect (1) the calcaneus (os calcis), a condition sometimes called apophysitis; (2) the capitular epiphysis (head) of the femur, a condition known as Legg-Calvé-Perthes disease, Perthes disease, Waldenstrom's disease, coxa plana, and pseudocoxalgia; (3) the ilium; (4) the lunate (semilunar) bone, known as Kienböck's disease; (5) head of the second metatarsal bone, known as Freiberg's infraction; (6) the navicular (tarsal scaphoid); (7) the tuberosity of the tibia, called Osgood-Schlatter disease and Schlatter's disease; (8) the vertebrae, called Scheuermann's disease or kyphosis, juvenile kyphosis, vertebral epiphysitis, and kyphosis dorsalis juvenilis; (9) the capitellum of the humerus, called Panner's disease.
The locations of the joints affected in human children can be contrasted with regions affected in the pig. In the pig, osteochondrosis can be located in the following areas, listed in descending order of severity of lesions: 1. Articular-epiphyseal lesions: stifle, elbow, lumbar synovial intervertebral joints, hock, shoulder, and hip, 2. Growth plate lesions: distal ulna, distal femur, costochondral junction, femoral head, humeral head, ischiatic tuberosity, and thoracolumbar vertebrae, 3. Epiphysiolysis and apophysiolysis lesions: glenoid cavity, ischiatic tuberosity, capital femoral epiphysis, vertebral epiphyses, anconeal process, and distal ulnar epiphysis.
Because of the similarity of the disease in pigs and humans, another embodiment of the invention is the treatment and prevention of OC in humans. Boron and vitamin C compounds can be administered to patients suffering from OC. The boron containing compounds useful for the practice of this invention may include any suitable organic, inorganic, or mineral boron containing compounds. Among the preferred forms of boron are sodium borate and boric acid. Other useful inorganic forms of boron include calcium borate. Examples of organic boron-containing compounds are well known to those of skill in the art. Examples of such organic boron-containing compounds are found in U.S. Pat. Nos. 4,312,989, 4,499,082, and 5,312,816. Dosages that may find use in humans include 1-13 ppm. Among the vitamin C compounds that may be used in the practice of the present invention are ascorbic acid, sodium ascorbate, calcium ascorbate, potassium ascorbate, and magnesium ascorbate.
Described below are administration methods that are useful for humans. One particularly useful administration method is the provision of boron and vitamin C as mineral or vitamin supplements, for example, in food or pill format. However, it will be appreciated that many of the methods disclosed below, while especially applicable to humans, can also be used for the administration of boron and vitamin C to animals as well.
One especially useful form of administration for the boron and vitamin C containing compounds of the present invention is as a mineral supplement with vitamins that can be taken orally as a pill or added to food. Multi-vitamin and mineral supplements are useful in the maintenance and improvement of health by insuring adequate intake of micronutrients that are needed for disease prevention and to compensate for nutritional deficiencies that result from factors as inadequate dietary intake of essential nutrients. Vitamin and mineral preparations are commonly administered as general nutritional supplements or to treat specific medical conditions. Accordingly, the supplemental boron containing compounds of the present invention can be administered as mineral supplements with vitamins such as vitamin A, vitamin C, vitamin D, vitamin E, vitamin K, vitamin B1, vitamin B2, niacinamide, vitamin B6, vitamin B12, biotin, pantothenic acid, carnitine, silicon, molybdenum, germanium iron, phosphorus, iodine, magnesium, zinc, selenium, copper, chromium, potassium, choline, lycopene, and co-enzyme Q-10. Examples of mineral supplement formulations to which supplemental boron containing compounds can be added can be found in U.S. Pat. Nos. 4,752,479, 5,869,084, and 6,361,800. Such supplements containing the boron and vitamin C compounds of the present invention can be administered as chewable vitamin pills, or as supplements that can be added to beverages, or as supplements that can be added to foods.
In practicing the method of the present invention, the boron and vitamin C compounds may be administered per se or as components of a pharmaceutically acceptable composition. When used in medicine, the form of the supplemental and vitamin C boron compounds should be both pharmacologically and pharmaceutically acceptable.
Thus, the present invention may be practiced with the boron and vitamin C compounds being provided in pharmaceutical formulations, both for veterinary and for human medical use, comprising the active agent (the boron compound) together with one or more pharmaceutically acceptable carriers thereof and optionally any other therapeutic ingredients. The carrier(s) must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the formulation and not unsuitably deleterious to the recipient thereof. The active agent is provided in an amount effective to achieve the desired pharmacological effect, as described above, and in a quantity appropriate to achieve the desired daily dose.
The formulations include those suitable for oral, rectal, topical, nasal, ophthalmic, or parenteral (including subcutaneous, intramuscular, and intravenous) administration. Formulations suitable for parenteral administration are preferred.
The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the active compound into association with a carrier which constitutes one or more accessory ingredients. In general, the formulations may be prepared by uniformly and intimately bringing the active compounds into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product into desired formulations.
Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets, tablets, or lozenges, each containing a predetermined amount of the active ingredient as a powder or in the form of granules; or as a suspension in an aqueous liquor or a non-aqueous liquid, such as a syrup, an elixir, an emulsion, or a draught.
A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine, with the active compound being in a free-flowing form such as a powder or granules which optionally is mixed with a binder, disintegrant, lubricant, inert diluent, surface active agent, or discharging agent. Molded tablets comprised of a mixture of the powdered active compound with a suitable carrier may be made by molding in a suitable machine.
One desirable formulation of the composition for administration is in a powdered form for dissolution or dilution with water or another suitable beverage or liquid before use. Alternatively, the composition can be contained in a ready to use form as part of a fortified beverage in liquid form. Also, boron and vitamin C containing compounds can be added to milk replacers. The composition can also be contained in a pudding with a custard or flan like texture or in the form of a bar suitable for ready consumption.
A syrup may be made by adding the active compound to a concentrated aqueous solution of a sugar, for example sucrose, to which may also be added any accessory ingredient(s). Such accessory ingredient(s) may include flavorings, suitable preservatives, agents to retard crystallization of the sugar, and agents to increase the solubility of any other ingredient, such as a polyhydroxy alcohol, for example glycerol or sorbitol.
Formulations suitable for parenteral administration conveniently comprise a sterile aqueous preparation of the active compound, which preferably is isotonic with the blood of the recipient (e.g., physiological saline solution).
Nasal spray formulations comprise purified aqueous solutions of the active compound with preservative agents and isotonic agents. Such formulations preferably are adjusted to a pH and isotonic state compatible with the nasal mucous membranes.
Formulations for rectal administration may be presented as a suppository with a suitable carrier such as cocoa butter, hydrogenated fats, or hydrogenated fatty carboxylic acids.
Topical formulations comprise the active compound dissolved or suspended in one or more media, such as mineral oil, petroleum, polyhydroxy alcohols, or other bases used for topical pharmaceutical formulations.
In addition to the aforementioned ingredients, the formulations of this invention may further include one or more accessory ingredient(s) selected from diluents, buffers, flavoring agents, binders, disintegrants, surface active agents, thickeners, lubricants, preservatives (including antioxidants), and the like.
The following examples further demonstrate several preferred embodiments of this invention. While the examples illustrate the invention, they are not intended to limit the invention. The patents cited herein are incorporated by reference in their entirety.
Three groups of 19 pigs, Duroc and Yorkshire pigs were randomly blocked by breed, litter and weight. The basal diet consisted of commercial corn-soy diet containing 10 ppm boron.
Test diet group B was fed a basal diet plus 25 mg/kg boron as sodium borate decahydrate (borax). Test diet group A was fed a basal diet plus 25 mg/kg boron as sodium borate decahydrate (borax) and 250 mg/kg ascorbic acid.
Pigs were weighed at the beginning of the study, 4 weeks later and every 3 weeks until the end of the study. Animals were scored for soundness on a 5-point scale at each weighing. (Five-point scale: 1=no soundness defects; 2=minor soundness issues but still sound enough for retention as breeding animal; 3=not sound enough for retention for breeding but still marketable; 4=unsound, likely to be rejected at slaughter; 5=severely lame, requiring euthanasia for humane reasons.) Grading was done by the caretaker and the investigator at all weighing times. An experienced independent treatment-blind rater also evaluated the soundness of each animal and the ratings he provided were compared with the ratings of the caretaker and investigator by use of Cohen's Kappa test.
The animals were housed in a modern curtain sided barn with deep straw bedding, with ad libitum access to feed and water. Waterers and feeders were on a concrete pad but the remainder of the flooring was deep straw over a ground limestone and sand base. Floor space allowances exceeded the recommendations of the Guide for the Care and Use of Agricultural Animals in Agricultural Research and Teaching, First Revised Edition, 1999. Federation of Animal Science Societies. Savoy, Ill.
Pigs in the study were fed a typical corn-soybean meal diet which contained a proprietary commercial supplement from the Moorman's company at the manufacturer's suggested inclusion rate. The basal diet (no boron supplemented) was analysed and found to contain boron at the rate of 10 ppm which is typical for a corn-soybean meal based diet.
Animals were observed twice daily by the caretaker.
In mid October one pig was observed to be severely lame and was euthanized and necropsied. Three additional pigs, one lame Duroc, a sound York and a sound Duroc (littermate to the lame pig) were euthanized and necropsied for observational purposes in mid-November.
At the end of the study the pigs were transported to a laboratory where they were euthanized and necropsied. Six pigs in each of group B and group A were put on the control diet for 7 days at the end of the study.
At necropsy, samples of liver, heart, kidney, fat, skeletal muscle, proximal tibia, blood, and the rostral snout were retained and frozen at −40° C. prior to chemical analysis. All joints of the axial skeleton were evaluated for the presence of gross lesions of osteochondrosis and graded on a 5 point scale. (5-point scale: 1=no gross abnormalities; 2=minor imperfections in articular conformation or articular reddening present but no cartilage erosions; 3=cartilage intact but surface irregularities of the cartilage are present; 4=fissuring or erosion of articular cartilage is obvious; 5=full thickness cartilage lesions or cartilage flaps, osteochondritis dissecans obviously present).
The proximal femur (femoral head) and articular surfaces and growth plates associated with the stifle, hock, shoulder, elbow and carpus were sectioned with a band saw and approx. 0.5 to 1.0 cm sections fixed in formalin. Bone sections were decalcified with formic acid/sodium citrate, embedded in paraffin and sectioned at 5 microns. Sections were deparaffinized according to standard procedures. Two sections were made from each joint and growth plate and stained with either hematoxylin/eosin (H&E) or with toluidine blue (pH 4)/fast green for evaluation of the articular cartilage, subchondral bone and growth plate.
Mean joint lesion scores, growth rate and soundness scores by treatment and by factors boron and ascorbate were compared using an analysis of variance and t-test as appropriate. Soundness scores were dichotomised into binary categorical variables for lameness (soundness score>2) and for absence of defects (soundness<2), and the binary variables analysed by chi-square and logistic regression.
Tissue specimens stained with H&E were examined microscopically for the presence of lesions of osteochondrosis. Toluidine Blue staining allowed assessment of retention or loss of proteoglycan from the extracellular matrix (ECM) of the cartilage. Each tissue section was scored by a treatment-blind board-certified pathologist.
Further, specific joints and structures are selected for more detailed histomorphometric analysis. Both epiphyseal and growth plate specimens were obtained from boron treated and untreated animals.
The results show that boron supplementation can be effective in reduction of the incidence of osteochondrosis-associated lameness in growing swine. Animals supplemented with boron had healthier joints than those receiving the basal diet with no supplemental boron (
In further microscopic analysis of the cartilage, one hundred and forty-one (141) sections of articular cartilage and growth plate have been prepared by formate decalcification and staining with H&E and Toluidine blue. The toluidine blue (TBlue) stain provided a semi-quantitative measure of sulfated glycosaminoglycan (sGAG) content. The consistently higher TBlue staining intensity among the boron supplemented group suggested a higher sGAG content in cartilage from boron supplemented pigs. (This effect of boron has been confirmed by a sGAG analysis procedure.) Lesions were evaluated by an experienced board certified pathologist specializing in porcine tissues. Lesions are classified into 2 factors. One factor comprising presence or absence of necrosis, infarction, hemorrhage or eosinophilic matrix streaks provides a measure of the structural integrity of the articular surface, while another factor comprising hyperplasia and abnormal differentiation provides a measure of the condition of the growth plate associated with growth plate widening. Growth plate lesions associated with widening were found in 57% of tissues from the unsupplemented group as compared with 19% of the tissues from pigs receiving 25 ppm boron. Articular cartilage lesions (necrosis, hemorrhage, infarction, or streaks) were found in 21% of tissues from unsupplemented pigs as compared with 4% of tissues from pigs receiving 25 ppm supplemental B. These data indicate that supplementation with boron can improve the structural integrity of the articular cartilage and the growth plate.
Anecdotal evidence from continuing experimental use of boron added to feed or drinking water indicates a consistent and sustained positive response in swine in a variety of production settings and genetics. This evidence is described in Example 6.
Boron nutrition is necessary to maintain the glycosaminoglycan concentration of cartilage at normal, healthy levels required for the cartilage to perform its function of resisting compression forces or to maintain the weight bearing ability of the cartilage.
Proteoglycans, a major component of healthy cartilage, draw and hold water which allows them to bear weight. We have measured the major subcomponent of proteoglycan, glycosaminoglycan (GAG) and have found that it is significantly reduced in non-boron supplemented pig cartilage tissue. Our data and the literature would support the statement that cartilage with low GAG levels fails to function much more readily than cartilage with higher levels of GAG.
Hock joints and elbow joints were obtained at necropsy from pigs in two feeding groups: one group having been fed a commercial ration supplemented with 50 mg B/kg feed and the other being fed only the standard commercial pig ration.
Cartilage plugs were harvested from the articular surfaces of 40 hock and elbow joints using a #5 (10.5 mm) cork borer. The samples were papain digested and analyzed for concentration of sulfated glycosaminoglycan (sGAG) using a spectrophotometric microplate analysis procedure adapted from Farndale, et al (1986), using a chondrotin sulfate standard. This method measures total sulfated glycosaminoglycan, most of which is chondrotin sulfate and keratan sulfate components of aggrecan. Aggrecan is the large aggregating proteoglycan of cartilage responsible for creating and maintaining the osmotic pressure of cartilage. Quanititation of sGAG concentration thus provides a measure of the weight-bearing and anti-friction properties of cartilage and is a measure of cartilage quality and durability.
The sGAG concentration (dry matter basis) in the cartilage of the pigs fed 50 ppm boron was 11050 ng/g as compared with 5045 ng/g among the unsupplemented pigs. This difference was significant at p<0.01. These data indicate that the chondroprotective properties of boron are mediated at least in part via mechanism that lead to increased sGAG levels in the cartilage.
It was observed that when sows were fed diets containing 50 ppm supplemental boron during the late gestation and early lactation period, piglet quality as assessed by uniformity, growth, and general thrift was improved, and pre-weaning piglet mortality was reduced. A preliminary pilot study confirmed these observations. Sows were fed a standard corn-soy diet. Half of the sows received an oral administration of a boron supplement to provide 1 mg boron per kg body weight. The other half did not receive any supplementation. Preliminary analysis of the data from the first 600 pigs indicated that the provision of boron to the gestating and lactating sows reduced pre-weaning mortality from 23% to 16% and increased piglet weight at 12 days of age from 8.0 pounds to 8.5 pounds, as compared to the non-supplemented groups.
To test the effect of boron on sows and their litters, a trial was established in a large commercial swine operation during an outbreak of porcine viral reproductive and respiratory disease (PRRSV). Boron was administered orally to a group of 51 sows individually housed in crates at the rate of 1 mg boron per pound BW per day beginning 1 week prior to farrowing and continuing until the piglets were weaned at 14 days of age. Their performance was compared to that of cohort of 50 sows of identical genetics and identical housing and husbandry conditions that did not receive boron. Boron treatment was supplied as a single daily dosing and was discontinued at weaning. All sows were fed a standard commercial sow diet.
There were no effects on litter size at birth or piglet birth weight. Piglets raised by sows consuming boron weighed 9.01 lbs at 12 days of age as compared with 8.32 lbs for the piglets raised by control sows (p=0.06). Piglet mortality in the boron treated group was 15.2% as compared with 20.3% among the controls (p=0.03). Sows that were fed boron returned to estrus an average of 1.6 days quicker than control sows (p=0.047). Boron treated sows were 1.2 times as likely as control sows to conceive (p=0.04). This result would be expected to have significant positive impact in a commercial pig raising operation.
It is postulated that borate exerts its beneficial effects in OCD prevention by modulation and stabilization of the extracellular matrix (ECM). In tissues like cartilage that possess an abundant ECM consisting of proteoglycan and collagen, the main effect of boron is likely mediated by a change in the mechanic (material) properties of the cartilage ECM. However many other tissues with important functions do also possess ECM components and extracellular receptors, the structure of which may be stabilized by boron cross-links which improves their functionality in cell to cell signaling, receptor functions, and adhesion functions. It is postulated that the effect of borate on reproduction is modulated by this sort of mechanism.
A 28-day feeding trial was conducted in a large commercial farm setting with 144 crossbred pigs of initial body weight of 24 kg. Pigs were randomly allocated to 24 pens of 6 pigs per pen in a thermoneutral controlled environment barn with steel grid flooring. Each pen was equipped with a single-hole feeder. Water was available free-choice from a nipple drinker. Pigs were fed a commercial pig diet containing 0.5% phosphorus plus 0 or 50 mg/kg Boron and a calcium level of either 0.5 or 0.65% in a 2×2 factorial design. Feces were collected on the last 3 days of the study from each pen and a pooled aliquot was dried and submitted for chemical analysis. Yttrium oxide was added to the diet at 0.05% and served as a marker for phosphorus digestibility. Pig growth and feed consumption was measured at the end of the study with the pen as the experimental unit. Data was analyzed for effects of boron and calcium by univariate and multivariate analysis of variance and t-tests.
Supplemental boron increased average daily gain, improved feed conversion ratio and phosphorus digestibility, and reduced fecal phosphorus excretion per unit of growth (p<0.05). (Table 2). Daily feed intake was not significantly modified by boron or calcium level (p>0.20). Decreasing calcium level improved feed conversion (p<0.05). There were no significant interactive effects of boron x calcium on feed conversion or phosphorus excretion (p>0.25).
The digestibility and fecal excretion of phosphorus is of significant concern to animal agriculture. Phosphorus is a costly dietary ingredient and the phosphorus in animal wastes is a potential environmental pollutant. In the present study, the phosphorus excretion per unit of production was reduced by 15% by adding boron to the diet. Boron also produced a significant effect on overall feed conversion ratio. It is expected that these effects of boron would have a significant impact in reducing environmental pollution as well as reducing production costs in animal agriculture
It was hypothesized that boron mediates its effect in the pig by quadrivalent crosslinking. 3-nitrophenylboronic acid (3-NPB) which is an avid blocker of boron crosslinks was administered orally to pigs of about 100 kg body weight at the rates of 0 and 1 grams in combination with supplemental boron at the rates of 0 and 50 ppm in feed. The 3-NPB was administered for 10 days. The pigs were evaluated for lameness daily and euthanatized on day 13 and the joints and other organs were examined.
All of the pigs that received 3-NPB but no supplemental boron developed clinical manifestations of OCD within 10 days, but only one in 5 of the pigs receiving supplemental boron developed lameness when 3-NPB was provided (Table 3). A chi-square analysis indicated a significant (p<0.05) effect of 3-NPB in inducing lameness and a significant effect of supplemental boron in preventing the lameness induced by 3-NPB.
Examination of shoulder, stifle, hock and elbow joints showed that 3-NPB treated pigs with no supplemental boron had a higher prevalence of osteochondrosis lesions and a more intense severity of lesions than other treatment groups, with the lowest prevalence of lesions and the lowest severity among those pigs receiving supplemental boron and no 3-NPB. Administration of 3-NPB along with boron resulted in a prevalence and severity of gross joint pathology similar to that observed in unsupplemented pigs. (
It was concluded that 3-NPB is acting as a competitive inhibitor of borate.
It is generally considered that the pig is the archetypic model species for osteochondrosis in mammals (see Reiland S. Osteochondrosis in the pig. Acta Radiol 1-118, 1975) The cascade of pathophysiologic events that culminate in clinical manifestations of osteochondrosis (OCD) in the pig are generally believed to be those events that occur in the other mammalian species that develop OCD, particularly the horse, dog, ruminants and humans. Since the pig is the model for OCD in other mammals and it has been demonstrated that boron is useful for prevention and treatment of OCD in the pig, it logically follows that boron should have a similar effect in other mammals and the effect should be mediated by a similar biochemical mechanism.
Therefore, it was concluded that administration of 3-NPB in species that are known to be susceptible to OCD but at a low prevalence, specifically cattle, horses, and dogs, should produce cartilage lesions indicating presence of a boron receptor moiety in cartilage.
Three healthy Holstein steer calves of average weight about 250 lbs and three healthy Quarter Horse fillies of average weight of about 500 lbs were administered 3-NPB at the rate of 10 mg/kg body weight per day. The 3-NPB was given to the calves by daily intraperitoneal injection, while the horses were administered the daily dose of 3-NPB mixed in feed. All animals consumed a standard ration of commercial feed and free-choice alfalfa-grass mixed hay.
Lameness was first observed in the calves at day 7 of the 3-NPB treatment. One calf was euthanatized at treatment day 14 and the other two at treatment day 21. Severe OCD lesions were visible in the hock and elbow joints of all calves with increasing severity noted with increasing time on 3-NPB.
Among the horses treated with oral 3-NPB, one fully showed clear signs of front leg lameness during exercise on treatment day 10 and was euthanatized on day 14. The other two horses were euthanatized on treatment day 28, at which time lameness was visible during exercise in one of the two horses. OCD lesions of varying degree were observed in the shoulder, elbow, fetlock and hock joints of all horses. Among the notable lesions were a 1 cm×1 cm necrotic lesion of the cartilage was found on the proximal articular surface of left front PI at day 14, a developing flap lesion on the distal tibia at day 28 and in another horse, profound thinning of cartilage and obvious cartilage wear lines were observed in the left front fetlock and left elbow at day 28.
In experiments with normal healthy crossbred hounds of age 10 weeks (body weight 8 kg), the administration of 3-NPB as a single daily dose of 10 mg per kg body weight resulted in visible foreleg lameness as early as 12 days. Necropsy revealed gross lesions of necrosis and hemorrhage in the distal ulnar growth plate. There was also evidence of cartilage erosion in the articular surfaces of the distal tibia and the proximal ulna. The gross lesions in the distal ulnar growth plates resembled the pathologic changes associated with osteochondrosis in swine.
It is concluded that the pig is a suitable model for OCD in both ruminant and non-ruminant animals, including carnivores, and that boron receptor sites exist in all mammalian species, and that supplemental boron is expected to be an effective preventative and remedy in all mammalian species.
Boron was added to the diet of sows at 1 mg/kg body weight as boric acid. There was no negative effect on reproduction or fertility. The sows appeared to have normal estrus activity and normal conception rate, with no negative effects on cyclic activity or on pigs born or on piglet viability. Three sows that were noticeably and seriously lame became perfectly sound. Among a group of 95 piglets the birth to weaning mortality was 2 piglets. The usual mortality for this farm was about 5 to 7%. These piglets remained on boron 50 ppm plus ascorbate 125 ppm. One pig was euthanized and examined at a weight of about 85 pounds. No abnormalities were observed in any of the joints. All bones of the appendicular skeleton were sectioned on the band saw. Bones had excellent mineralization and the growth plates were narrow and crisply demarcated, including the distal ulnar growth plate which is an early predilection site for OC-related abnormalities.
An Iowa farm treated a group of about 100 pigs with 50 ppm boron as boric acid. None of these pigs developed any signs of lameness or unsoundness. The farmer reported that these pigs are the most sound he has raised. The estimated previous lameness/unsoundness rate was about 25 to 30%, and zero in the test group. The pigs demonstrated excellent growth rate. The absence of lameness and hock swelling was observed. Two pigs with hock swelling were euthanised from among the younger and older pigs not treated with boric acid. The younger pig of about 50 pounds body weight showed evidence of early OC changes in the hock. An older pig of about 250 pounds body weight with severe lameness in the right hock was euthanised. Severe advanced OCD was observed in the hock, and growth plate abnormalities were observed when the bones were sectioned. Culture of the hock joints was negative, ruling out bacterial infection and indicating that OC is the likely cause of lameness.
One hundred commercial hybrid tom turkeys at age 13 weeks were selected randomly from a healthy flock of 4700 birds and were randomly allocated to two treatment groups (50 birds per group) for a trial of the effects of boron and ascorbate on leg structure and gait abnormalities. Many of the birds were large enough to be showing early signs of leg weakness such as trembling and gait abnormalities. Birds were evenly distributed between treatments on signs of leg weakness.
One treatment group (“Control”) was fed the standard commercial dietary regimen and plain drinking water, while another group (“B+C Treatment”) was fed the standard commercial diet but received supplemental boron and supplemental ascorbic acid in the water. The rate of boron and ascorbic acid in the water was adjusted to provide supplementation similar to the rate that would be provided by an in-feed supplementation of 50 mg of elemental boron and 50 mg of ascorbic acid per kilogram (50 ppm for boron and 100 ppm vitamin C) of dry feed. The trial was started on the 12th of August of year 1 and the birds were euthanatized and necropsied on September 12, of year 1. Mortality and morbidity loss and weight gain was similar between the two treatment groups. Mortality and morbidity losses resulted in 92 birds remaining at the end of the trial.
The birds were scored for lameness/mobility/gait abnormality on a five point scale as follows: Grade 1=good mobility without trembling or hesitation. Grade 2=good mobility but some trembling or hesitation is observed. Grade 3=definite trembling when standing, and definite reluctance to move about. Grade 4=marked reluctance to walk, bird is recumbent and generally rises to walk only when forced to move. Grade 5=bird is either unable or nearly unable to walk.
The femur and tibia were dissected from each leg of each bird, and the proximal tibia was transected by sawing in the transverse plane for evaluation of the proximal growth plate.
Growth plates were measured with a digital caliper and the widest portion of the growth plate among the sections of the right and left tibias was recorded. Large cartilage plugs typical of classical tibial dyschondroplasia were not included in the growth plate measure. Tibial dyschondroplasia was evaluated by the presence or absence of the typical triangular cartilage plug in the proximal tibia, and a subjective “td score” was assigned based on the size of lesion, with 0 indicating no cartilage plug, and 3 the most severe form. Cartilage plugs were considered to be “active” if they were confluent with the growth plate and “old” or “healed” if not confluent with the growth plate but were rather separated from the growth plate by layers of bone. The presence of overt grossly observable necrosis and haemorrhage growth plate was graded from 0 to 3 with 0 being no changes observed and 3 representing severe necrosis and haemorrhage with detachment of the proximal epiphysis of the tibia.
The data from subjective and objective measures were analyzed by parametric and non-parametric statistical methods and appears in Table 4.
1Maximum width of proximal tibial growth plates including right and left, not including classical “TD cartilage plugs”.
2NH scoring on the presence and severity of necrosis and haemorrhage in the growth plate, right and left collectively
3Cartilage plugs observed in active form in either right or left proximal tibia.
4Scoring of active and “healing” cartilage plugs (subjective).
5Subjective score including only active cartilage plugs confluent with growth plate.
One hundred tom turkeys were selected at random at 9 weeks of age and divided into two groups, control and B+C (boron and ascorbate supplemented, as described above) of 50. Each bird was weighed and tagged at week 9. At 17 weeks, all birds were weighed, scored for lameness/mobility/gait abnormality using the five point scale used in study 1, and sacrificed. The tibia was dissected from each leg of each bird and subjected to an ultimate breaking force analysis (3-point bending test).
Lameness/mobility/gait abnormality results were substantially similar to study 1, showing improvement for the B+C group, as compared to control.
250 turkeys were selected at random at week 11 and divided into 5 treatment groups: 1) control, 2) ascorbate only “(VC”), 3) boron only (“B”), 4), boron and ascorbate (“B+C”), and 5) ½ rate boron and full rate ascorbate (“LoB+C”). Full rates of both boron and ascorbate were as described for study 1. At 17 weeks, all birds were weighed, scored for lameness/mobility/gait abnormality using the five point scale used in study 1, and sacrificed. The femur was dissected from each leg of each bird and subjected to an ultimate breaking force analysis. Cartilage at the femoral distal condyle was subjected to an indenter test.
For the lameness/mobility/gait abnormality scoring, three graders were used. Two of the graders were “blind” graders unaware of each turkey's grouping. The third grader was a “biased” grader aware of each turkey's grouping. Partial grading on the 5 point scale described above was allowed.
Overall, a combination of boron and ascorbate administered in drinking water was found to reduce the incidence of leg weakness and severity of leg weakness scores in turkeys, and improved bone strength as measured by a 3-point bending test. The administration of ascorbate alone improved cartilage strength as measured by an indenter test, and slightly improved bone strength as assessed by a classic 3-point bending test of the femur, but ascorbate alone did not significantly improve leg weakness scores. The administration of boron alone improved bone strength by 7.8% as compared with the control, and reduced leg weakness scores to a mean of 2.3 as compared with 3.0 in the control (5 point scale). Growth rates were improved when boron was provided. The administration of boron and ascorbate in combination improved leg weakness scores to a mean of 1.5 as compared with 3.0 in the control and 2.3 in the birds treated with boron alone. This demonstrates an interactive effect of the combination of boron and ascorbate, since ascorbate alone did not significantly change leg scores but ascorbate did result in a significant incremental reduction in leg weakness when added in combination with boron.
Assuming similar results on commercial scale-up, it may be possible that the post-12 week mortality due to leg weakness will be reduced by 50 to 90% for the application of the boron and ascorbate treatment, for a potential production cost reduction extrapolated at $100 million to $180 million for the U.S. national turkey industry.
This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 60/873,420, filed Dec. 6, 2006, which is hereby incorporated by reference, in its entirety.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/US07/25103 | 12/6/2007 | WO | 00 | 6/4/2009 |
| Number | Date | Country | |
|---|---|---|---|
| 60873420 | Dec 2006 | US |
