Methods for: reducing the risk of peripartum hypocalcemia in at least one ruminant in the periparturious period, and assessing whether a ruminant in the periparturious period is susceptible to a treatment to reduce the risk of periparturior hypocalcemia, composition for reducing the risk of periparturious hypocalcemia, its use, and use of a phosphorus chealant compound

Description

1 / 19 “METHODS FOR: REDUCING THE RISK OF PERIPARTUM HYPOCALCEMIA IN AT LEAST ONE RUMINANT ANIMAL IN THE PERIPARTURIENT PERIOD, AND EVALUATING WHETHER A RUMINANT IN THE PERIPARTURIENT PERIOD IS SUSCEPTIBLE TO TREATMENT TO REDUCE THE RISK OF PERIPARTUM HYPOCALCEMIA, COMPOSITION TO REDUCE THE RISK OF PERIPARTUM HYPOCALCEMIA, ITS USE, AND USE OF A PHOSPHORUS CHELATING COMPOUND” CROSS-REFERENCED TO RELATED APPLICATIONS

[001] This PCT application claims priority from U.S. Non-Provisional Patent Application No. 18 / 830,440, entitled COMPOSITION AND METHOD FOR REDUCING THE RISK OF HYPOCALCEMIA IN PERIPARTURIMENTAL RUMINANT ANIMALS, filed September 10, 2024, which is a partial continuation of U.S. Non-Provisional Patent Application No. 18 / 618,702, entitled COMPOSITION AND METHOD FOR REDUCING THE RISK OF HYPOCALCEMIA IN PERIPARTURIMENTAL RUMINANT ANIMALS, filed March 27, 2024, which claims priority from U.S. Provisional Patent Application No. 63 / 601,401, entitled COMPOSITION AND METHOD FOR REDUCING THE RISK OF HYPOCALCEMIA IN ANIMALS PERIPARTURIMENTAL RUMINANTS, filed on November 21, 2024. 2023, the contents of which are incorporated herein by reference in their entirety. FIELD

[002] This application relates to the prevention or treatment of hypocalcemia in periparturient ruminant animals. BACKGROUND

[003] Milk fever (hypocalcemia) in dairy cows (and other lactating animals) is a metabolic disorder that occurs around the time of calving. Severe clinical cases are characterized by levels lRutçth>87(E!60053ffi5!, dte03 / (W>,2ffi!6, pág. 45 / 118 2 / 19 very low blood calcium levels, resulting in cows that are lethargic and unresponsive to stimuli. Milk fever can be classified as Clinical Hypocalcemia (blood calcium less than 7.0 mg / dl) or Subclinical Hypocalcemia (blood calcium between 7.0 and 8.5 mg / dl). Clinical hypocalcemia (CH) affects 3 to 6% of all cows in their second lactation or older. Subclinical hypocalcemia affects 60 to 80% of cows in their second lactation or older. Subclinical hypocalcemia (SCH) may not present physical symptoms, but borderline blood calcium levels during the first 4 days postpartum have been shown to be a significant risk factor for metabolic diseases. Research shows that cows with subclinical hypocalcemia (SCH) have a higher risk of ketosis, retained placenta, metritis, displaced abomasum, and mastitis compared to those with normal blood calcium levels (> 8.5 mg / dl).Although the symptoms of SCH are not as severe as those of chronic hypocalcemia (CH), the fact that it affects a very high percentage of cows in the peripartum period and still represents a health risk suggests that SCH can be as costly, or even more so, to a dairy herd as CH.

[004] Hypocalcemia is caused by the high calcium demand at calving for colostrum synthesis. Colostrum has twice the calcium concentration compared to regular milk. When colostrum synthesis begins, the cow's homeostatic mechanism in the peripartum period is unable to maintain normal blood calcium levels, which is especially true for second-lactation and older dairy cows. Cows in the peripartum period have a negative calcium balance, having 8 to 9 grams of body calcium available, but needing up to 30 grams of calcium per day for colostrum synthesis. The calcium deficit needs to be supplied by the bones to maintain normal blood calcium levels and lRutçth>87(E!600538SB, dte03 / (W>.affi!6,|piig. 46 / 118 3 / 19 Cow health. Research has focused on trying to elucidate the metabolic reasons for the cow's inability to maintain blood calcium levels.

[005] Dietary mineral binders, such as calcium binders, are a known strategy in feeding cows during the peripartum period. By binding to a mineral, such as dietary calcium, a real calcium deficiency is induced, resulting in the stimulation of parathyroid hormone and the activation of vitamin D, which allows the cow to better respond to the demand for calcium in the blood at the time of calving. Essentially, calcium binders ensure that the blood's homeostatic mechanism is functioning efficiently at the time of calving, when it is most needed. In addition, mineral binding beyond calcium can be done, with feeding with a phosphorus binder being a relatively new strategy that has been used in the sector.Through the binding of phosphorus, using an aluminum-based product, a reduction in blood phosphorus occurs, stimulating the homeostatic mechanisms necessary for the cow in the peripartum period to respond to the challenge of calcium demand. This product is provided, but optimizing the supply rate and the amount of dietary phosphorus to be bound relative to aluminum can be a challenge.

[006] Furthermore, the dietary phosphorus to aluminum ratios in total mixed rations (TMR) used in feeding plans on dairy farms in the United States are inconsistent, varying considerably from farm to farm when these aluminum-based products that bind to phosphorus are provided. Therefore, there is a lack of precision in the P:Al ratios in the feeds being used. Thus, there is a need for a method in which the P:Al ratio can be consciously adjusted with the aim of maintaining blood phosphorus concentration within a desired level for lRutçã(>87(E!60053ÍSB, dte03 / (W>.affi!6, pág. 47118 4 / 19 helps reduce the risk of peripartum hypocalcemia in dairy cows.

[007] Therefore, there remains a need for effective strategies to reduce the risk of hypocalcemia in the peripartum period of ruminant animals, especially cows. SUMMARY

[008] The present invention overcomes several deficiencies, disadvantages and undesirable parameters associated with known methods and compositions for reducing the risk of periparturient hypocalcemia in periparturient ruminant animals.

[009] In one embodiment, the present invention relates to a method for reducing the risk of periparturient hypocalcemia in periparturient ruminant animals. The method comprises the steps of a) measuring the concentration of phosphorus (P) in the animal's blood; and b) administering to the animal, during at least part of the prepartum period, the composition comprising a phosphorus-binding compound and, optionally, a pharmacologically acceptable vehicle. The composition comprises an effective amount of the phosphorus-binding compound to control the concentration of phosphorus in the blood in a range of 1.5 to 3.5 mg of P per dL of blood in the periparturient ruminant animal.

[0010] In a second embodiment, the present invention relates to a composition for reducing the risk of periparturient hypocalcemia in periparturient ruminant animals. The composition comprises a phosphorus-binding compound present in an amount that controls the phosphorus concentration in the blood of the periparturient ruminant animal in a range of 1.5 to 3.5 mg P per dL of blood; and, optionally, a pharmacologically acceptable carrier.

[0011] Optionally, the blood phosphorus concentration from step a) is controlled to be in a range of 1.8 to 3.0 mg P per dL of blood. lRutçth>87(E!600538SB, Jte03 / (W>,2ffi!6, p. 48 / 118 5 / 19

[0012] Optionally, the blood phosphorus concentration from step a) is controlled to be in a range of 2.0 to 2.5 mg P per dL of blood.

[0013] If the blood phosphorus concentration is below the range, the administration rate of the compound in step b) is decreased or the compound is reformulated to decrease the amount of the compound and / or increase the phosphorus to aluminum ratio, so that the blood phosphorus concentration is brought into the range.

[0014] If the blood phosphorus concentration is above the range, the administration rate of the compound in step b) is increased or the compound is reformulated to increase the amount of the compound relative to phosphorus in the compound and / or decrease the phosphorus-to-aluminum ratio, so that the blood phosphorus concentration is brought into the range.

[0015] Optionally, blood phosphorus levels are measured periodically.

[0016] Preferably, the phosphorus-binding compound in the composition reduces the bioavailability of phosphorus in the digestive system of the periparturient ruminant animal.

[0017] Optionally, the periparturient ruminant animal is a dairy cow.

[0018] Optionally, the composition is an animal feed composition, and the pharmacologically acceptable vehicle comprises one or more animal feed components other than the phosphorus binding compound.

[0019] Optionally, the phosphorus linking compound comprises an aluminum compound.

[0020] Optionally, the animal feed composition provides a phosphorus (P) to aluminum (Al) weight ratio in the diet in a range of 0.7:1 to 1.4:1. lPUçth>87(E!60053ffiB, dte03 / (W>,2ffi!6, p. 49 / 118 6 / 19

[0021] Optionally, the weight ratio of phosphorus to aluminum in the diet is in a range of 0.8:1 to 1.1:1.

[0022] Optionally, the aluminum compound comprises an aluminosilicate, aluminum sulfate, aluminum chloride, aluminum ammonium sulfate, aluminum potassium sulfate, aluminum sodium sulfate, aluminum calcium silicate, aluminum hydroxide, aluminum bromide, aluminum iodide, aluminum nitrate, aluminum carbonate, a hydrate thereof, or any mixture thereof.

[0023] Optionally, the aluminum compound comprises aluminum sulfate, aluminum chloride, aluminum hydroxide, a hydrate thereof, or any mixture thereof. DESIGNS

[0024] These and other features, aspects and advantages of the present invention will be better understood with reference to the following description, the appended claims and the accompanying drawings.

[0025] Fig. 1 presents a flowchart of a first method for reducing the risk of periparturient hypocalcemia in a periparturient ruminant animal, exhibiting features of the present invention; Figure 2 presents a graph showing the relationship between blood phosphorus (P) (mg / dL) and the P:Al ratio (m / m) in the diet, discussed in Example 1; and Figure 3 presents a flowchart of a second method for reducing the risk of periparturient hypocalcemia in a periparturient ruminant animal, exhibiting characteristics of the present invention. DESCRIPTION

[0026] As used herein, the following terms and their variations have the meanings assigned below, unless a different meaning is clearly intended by the context in which such term is used. lPeitçth>87(E!60053ffi5!, dte03 / (W>,2ffi!6, pg. 50 / 118 7 / 19

[0027] The terms a, an, and similar referents used here are to be constructed to cover both the singular and the plural, unless their use in the context indicates otherwise.

[0028] As used in this description, the term "comprise" and variations of the term, such as "comprising" and "comprises," are not intended to exclude other additives, components, ingredients, or steps.

[0029] The term birth refers to the period during which an animal gives birth to a young animal.

[0030] The term pre-partum refers to the period before childbirth.

[0031] The term postpartum refers to the period following childbirth.

[0032] The term peripartum refers to the period of childbirth and the events that occur a few weeks before and after childbirth.

[0033] Referring now to Fig; 1, a flowchart of a method for reducing the risk of peripartum hypocalcemia in a ruminant animal is presented.

[0034] There is a strong correlation between blood phosphorus and calcium levels. Therefore, the concentration of phosphorus in the blood of a periparturient ruminant animal is related to the risk of hypocalcemia. It is also believed that the amount of phosphorus in animal feed influences the incidence of hypocalcemia in periparturient ruminant animals. By reducing the bioavailability of phosphorus in the digestive system, especially in the small intestine, during at least part of the prepartum period, it is possible to reduce the risk of peripartum hypocalcemia. The reduction of phosphorus bioavailability in the digestive system is preferentially achieved by binding phosphorus in the diet.

[0035] Phosphorus-binding compounds form complexes or compounds with dietary and gastrointestinal phosphorus, thereby reducing phosphorus absorption. Phosphorus-binding compounds are therefore particularly useful in reducing the risk of hypocalcemia. 8 / 19 periparturient in the animal, whether the compound is administered independently of animal feed rations or as a component of animal feed rations.

[0036] The phosphorus-binding compound preferably has a phosphorus-binding capacity of at least 60 mg of phosphorus per gram of phosphorus-binding compound, more preferably at least 65 mg of phosphorus per gram of phosphorus-binding compound.

[0037] The binding capacity may be in the range of 60–130 mg of phosphorus per gram of phosphorus-binding compound. Certain pharmacologically acceptable aluminum compounds have been found to be very effective as phosphorus-binding compounds. In some embodiments, the aluminum compound comprises an aluminosilicate (e.g., a zeolite), aluminum sulfate, aluminum chloride, aluminum ammonium sulfate, aluminum potassium sulfate, aluminum sodium sulfate, aluminum calcium silicate, aluminum hydroxide, aluminum bromide, aluminum iodide, aluminum nitrate, aluminum carbonate, a hydrate thereof, or any mixture thereof.The aluminum compound preferably comprises aluminum sulfate, aluminum chloride, aluminum hydroxide, a hydrate thereof, or any mixture thereof, for example, aluminum sulfate, aluminum hydroxide, a hydrate thereof, or any mixture thereof. In some embodiments, the aluminum compound is not an aluminosilicate.

[0038] The phosphorus binding compound is used in a pharmacologically acceptable amount that is effective in reducing phosphorus absorption by the animal from drinking water and / or feed. In some embodiments, the phosphorus binding compound is used in the composition of animal feed, for example, a total mixed ration (TMR). In some embodiments, the phosphorus binder comprises an aluminum compound, and the amount of aluminum compound lRutçã(>87(E!60053ffiB, dte03 / (W>,2ffi!6,|piig. 50 / 118 The 9 / 19 ratio used in the feed composition provides a phosphorus (P) to aluminum (Al) weight ratio in the diet ranging from 0.7:1 to 1.4:1. In some formulations, the weight ratio is 0.8:1 to 1.2:1. In some formulations, the weight ratio is 0.8:1 to 1.1:1. In some formulations, the weight ratio is 0.9:1 to 1.1:1.

[0039] Reducing the risk of hypocalcemia is most effective when the concentration of phosphorus (P) in the animal's blood is maintained within a specific range. In some embodiments, the concentration of phosphorus in the animal's blood is controlled within a range of 1.5 to 3.5 mg P per dL of blood. In some embodiments, the concentration of phosphorus in the blood is within a range of 1.8 to 3.3 mg P per dL of blood. In some embodiments, the concentration of phosphorus in the blood is within a range of 1.8 to 3.0 mg P per dL of blood. In some embodiments, the concentration of phosphorus in the blood is within a range of 2.0 to 3.0 mg P per dL of blood. In some embodiments, the concentration of phosphorus in the blood is within a range of 2.0 to 2.5 mg P per dL of blood. In some embodiments, the phosphorus-binding compound is used in a P:Al ratio in the range of 0.7:1 to 1.4:1 w / w to control the concentration of phosphorus in the blood in a range of 1.5 to 3.5 mg of P per dL of blood in the animal.In other words, in a pre-partum cow, the concentration of phosphorus in the blood is in the range of 2.0 to 3.0 mg / dL. In a recently calved cow (preferably less than 6 hours after calving), the concentration of phosphorus in the blood is in the range of 1.8 to 3.0 mg / dL. In a post-partum cow (preferably more than 2 days after calving), the concentration of phosphorus in the blood is in the range of 3.5 to 6.0 mg / dL.

[0040] Maintaining blood phosphorus concentration within these ranges has the most profound positive effect on reducing the risk of hypocalcemia. For this reason, measuring phosphorus concentration lRutçth>87(E!60053ffiB, dte03 / (W>,2ffi!6, pág. 53118 A 10 / 19 phosphorus level in the blood is useful. Measuring blood phosphorus concentration is done using known methods. In some methods, the measurement is taken periodically over a period of time, for example, once or twice or more per week. In some methods, measuring blood phosphorus concentration involves collecting a blood sample from the animal.

[0041] If the blood phosphorus concentration is below the desired range, the administration rate of the composition may be decreased or the composition may be reformulated to decrease the amount of phosphorus-binding compound relative to phosphorus in the composition. If the blood phosphorus concentration is above the desired range, the administration rate of the composition may be increased or the composition may be reformulated to increase the amount of phosphorus-binding compound relative to phosphorus in the composition. In either case, the blood phosphorus concentration is preferably brought to the desired range.

[0042] The phosphorus-binding compound or the composition comprising it is preferably formulated for oral administration. The composition may be formulated with the phosphorus-binding compound alone or with a pharmacologically acceptable carrier.

[0043] Consequently, the method of the present invention relates to a method for reducing the risk of periparturient hypocalcemia in periparturient ruminant animals. The method comprises the steps of: a) measuring the concentration of phosphorus (P) in the blood of the periparturient ruminant animal; and b) administering to the periparturient ruminant animal, during at least part of the preparturient period, a composition. The composition comprises an effective amount of a phosphorus-binding compound to control the concentration of phosphorus in the blood in a range of 1.5 to 3.5 mg of P per dL of blood in the periparturient ruminant animal; and, optionally, a vehicle. 11 / 19 pharmacologically acceptable.

[0044] If the blood phosphorus concentration is below the range, the administration rate of the compound in step b) is reduced or the compound is reformulated to decrease the amount of the compound and / or increase the phosphorus to aluminum ratio.

[0045] If the concentration of phosphorus in the blood is above the range, the administration rate of the composition in step b) is increased or the composition is reformulated to increase the amount of the compound relative to phosphorus in the composition and / or decrease the phosphorus to aluminum ratio.

[0046] Optionally, the blood phosphorus concentration measurement used to adjust the administration of the composition or to reformulate it is an average blood phosphorus concentration obtained from a plurality of animals.

[0047] In some embodiments, the composition may be formulated as an orally administerable solution, preferably an aqueous solution, or as a dry formulation (e.g., a powder, a tablet, a capsule, or similar) and administered separately or concomitantly with the feed.

[0048] Preferably, the composition is a feed composition, for example, total mixed ration (TMR), wherein the phosphorus-binding compound is formulated together with one or more animal feed components other than the phosphorus-binding compound. One or more animal feed components are the pharmacologically acceptable carrier and may include, for example, one or more of hay, vegetable protein products (e.g., soy protein, corn gluten meal (FGM)), grain products (e.g., barley, rye, oats, wheat straw and the like), processed grain by-products (e.g., grain silage), processed corn by-products (e.g., corn silage, corn starch), molasses (e.g., lPeitçã(>87(E!60053ffiB, dte03 / (W>,2ffi!6, pág. 55 / 118 12 / 19 condensed molasses, isomaltulose), fatty acids (e.g., conjugated linoleic acid (CLA)), mineral supplements (e.g., calcium carbonate, monocalcium phosphate, sodium chloride, magnesium oxide, copper sulfate, calcium sulfate, manganese sulfate, zinc sulfate, potassium chloride, potassium sulfate, magnesium sulfate, cobalt carbonate, sodium selenite, magnesium (mica)), vitamin supplements (e.g., vitamin E, activated animal sterol with D3 (a source of vitamin D3), vitamin A), ethylenediamine dihydroiodide, metal-amino acid complexes (e.g., zinc amino acid complex, manganese amino acid complex, copper amino acid complex), mineral oil, preservatives (e.g., propionic acid), dried brewer's yeast, plant extracts (e.g., yucca schidigera extract, fenugreek flavor extract, hemicellulose extract), betaine hydrochloride, acid Lactic acid, whey powder,Beneficial bacteria (e.g., Bactilus subtilis fermentation product powder, Bifidobacterium longum fermentation product powder, Bifidobacterium thermophilum fermentation product powder, Enterococcus faecium fermentation product powder, Lactobacillus acidophilus fermentation product powder), seaweed flour powder (e.g., Fucaceae, Bangliceae, Ulvaceae), psyllium seed husk, chicory root, red pepper, vegetable oils (e.g., glycerin, soybean oil, anise oil, oregano oil, lemongrass oil), cloves, natural flavor, and silicon dioxide, among others.

[0049] In feed compositions, the phosphorus binding compound is preferably present in an amount of 3% by weight or less of the phosphorus binding compound, based on the total dry matter intake of the feed consumed by the animal, preferably from 0.1 to 3% by weight, for example, about 1 to 2% by weight.

[0050] The animal is a ruminant, for example, a cow, a sheep, lRutçth>87(E!60063ffiB, dte03 / (W>,2ffi!6,|piig. 56118 13 / 19 a goat or similar. In a preferred embodiment, the animal is a cow, for example, a dairy cow or a beef cow. The animal is preferably a dairy cow. The ration composition is preferably formulated in a manner suitable for a cow.

[0051] Referring now to Figure 3, a second method is shown for reducing the risk of periparturient hypocalcemia in a periparturient ruminant animal, exhibiting features of the present invention. The method comprises the steps of measuring the aluminum in the diet (the amount / concentration of aluminum in the ration composition) and adjusting the amount / concentration of aluminum in the diet to ensure that the animal's blood phosphorus level is within the ideal range (Pre-partum cows: 2.0-3.0 mg / dL, Newly calved cows (< 6 hours): 1.8-3.0 mg / dL and Post-partum cows (> 2 days): 3.5-6.0 mg / dL).

[0052] More specifically, ideal levels of aluminum in the diet are between 0.25-0.40% (or 2500-4000 ppm) (determined by means of a wet chemical analysis of the Total Mixed Ration). Preferably, levels of aluminum in the diet are between 0.28-0.39% (or 2800-3900 ppm). These levels of aluminum in the diet relative to phosphorus in the diet result in the desired ratio of 0.80-1.20 of phosphorus to aluminum in the diet, cited above in paragraph

[0040] .

[0053] If the aluminum concentration in the feed composition is below one of the ranges mentioned above (below 0.25% or 2500 ppm) (determined by wet chemical analysis of the Total Mixed Ration - TMR), the feeding rate of the feed composition is increased or the feed composition is reformulated to increase the amount of aluminum in the feed composition.

[0054] If the concentration of aluminum in the feed composition exceeds one of the ranges mentioned above (above 0.40% or 4000 ppm), the feeding rate of the feed composition is reduced or the feed composition is reformulated to decrease the amount of lRutçth>87(E!60053ÍSB, dte03 / (W>,2ffi!6, pág. 57 / 118 14 / 19 aluminum in the feed composition. EXAMPLES Example 1: Method for Feeding a Product Containing Aluminum Using the P:Al Ratio in the Diet

[0055] To optimize the feeding rate of an aluminum-containing product (e.g., zeolite A), a dietary P:Al weight ratio was developed that achieves the desired reduction in blood phosphorus concentration. In the method, a P:Al ratio of 0.8:1 to 1.1:1 reduces blood P to approximately 1.8–3 mg / dL and requires three methodological steps: (1) formulation and feeding of the diet to achieve a dietary P:Al ratio of 0.8:1 to 1.1:1, (2) confirmatory tests of feed samples to validate and confirm the ideal dietary P:Al ratio, and (3) analysis and confirmation of the reduction in blood P concentrations between 1.8–3 mg / dL in peripartum cows fed the Al-containing product, which was optimized for a dietary P:Al ratio of 0.8:1 to 1.1:1. Step 1. Diet Formulation and Feeding

[0056] To achieve the ultimate goal of restricting dietary and blood P using an Al-containing product, a dietary P:Al ratio of 0.8:1 to 1.1:1 (Fig. 2) was formulated and provided. In a pre-partum dairy cow diet, the basal aluminum (Al) content is approximately 400 ppm on a dry basis, or a dietary P:Al ratio of 8:1 to 10:1, when grains, protein sources, and silages are mixed to form a total mixed ration (TMR). Thus, to achieve a dietary P:Al ratio of 0.8:1 to 1.1:1, an aluminum-containing product is supplemented in the TMR to increase the Al content and allow the formation of the aluminum phosphate complex in the animal's gastrointestinal tract, reducing the available dietary P. The supplementation rate of the product containing Al is adjusted according to dietary P levels to achieve the target range of the P:Al ratio mentioned above. 15 / 19 (0.8:1 to 1.1:1). As the P content in the diet (in % of mass) increases, Al supplementation also increases. Consequently, as the P content (in % of weight) in the diet decreases, supplementation with an Al-containing product is reduced to maintain a P:Al ratio in the diet of 0.8:1 to 1.1:1, based on the feed ingredients. Step 2. Diet confirmation

[0057] Variations in dietary P and Al levels can occur due to feed formulation, feeding, and animal availability, for various reasons. Therefore, confirmation of the formulation and feeding is performed to verify if the P:Al ratio of 0.8:1 to 1.1:1 is being met in the actual diet. To measure the P and Al content in the feed and thus determine the dietary P:Al ratio, at least two samples of the total mixed ration (TMR) provided to cows pre-partum were collected. After collecting the TMR samples from the feed bunk or mixer wagon, the samples were sent to a forage analysis laboratory to be analyzed for P and Al content using inductively coupled plasma optical emission spectrometry (ICP). ICP analysis is an analytical technique capable of identifying and quantifying the amount of P and Al in a sample based on the ionization of the elements in response to intense heating.The spectral emissions from ionization are then quantified in a spectrometer, which translates into concentrations of specific elements. It is important that the analysis of P and Al be performed by ICP due to the greater linear range, thus ensuring the accuracy of the measurements.

[0058] Once analyzed, the P and Al content in the diet is available on a dry matter basis to calculate the P:Al ratio in the diet and confirm whether the diet formulation is aligned with the actual feed mixture. If the P:Al ratio in the diet is within the range of 0.8:1 to 1.1:1, the methodology advances to the step of confirming P in the blood. If the lRutçth>87(E!60053ffiB, dte03 / (W>.affi!6, pág. 59 / 118. If the 16 / 19 P:Al ratio in the diet is above the target (greater than 1.1:1), adjustments to the formulation were made. These adjustments can be made by increasing the Al-containing product to decrease the P:Al ratio in the diet or by decreasing the P in the diet through feeding with alternative forages or protein sources to decrease the P:Al ratio in the diet and keep it within the target range of 0.8:1 to 1.1:1. If the P:Al ratio in the diet is too low (less than 0.8:1), the diet formulation was also adjusted. By decreasing the Al-containing product in the formulation or increasing the P in the diet, the goal was to increase the P:Al ratio in the diet so that it was within the desired range of 0.8:1 to 1.1:1. After adjustments to the formulation and the collection and analysis of new feed samples, and with the P:Al ratio in the diet between 0.8:1 and 1.1:1, the methodology advances to confirm whether adequate blood P levels have been achieved. Step 3. Confirmation of Blood Phosphorus

[0059] A dietary P:Al ratio between 0.8:1 and 1.1:1, based on the data presented here (Fig. 2), results in a blood P concentration of 1.8 to 3 mg / dL. To confirm whether optimizing the dietary P:Al ratio resulted in a reduction of blood P to the mentioned range, blood samples were collected from cows during the peripartum period and / or immediately after calving as follows. Blood Sampling in the Prenatal Period

[0060] Blood samples were collected from at least ten dairy cows in the pre-partum period after consuming a diet supplemented with an aluminum-containing product (zeolite A) to achieve a dietary P:Al ratio of 0.8:1 to 1.1:1. The cows consumed the Total Mixed Ration (TMR) supplemented with the aluminum-containing product for at least 7 days before being eligible for blood collection. Blood samples were collected from the coccygeal vessel at the base of the tail and analyzed to determine blood phosphorus concentrations. lRutçth>87(E!60053ffiB, de07SO7 / 2026,,pág;. 60118 17 / 19 Postpartum Blood Sampling of Cows

[0061] At least ten dairy cows were collected from blood samples between 6 and 12 hours postpartum, after consuming a diet supplemented with the aluminum-containing product, formulated and analyzed to achieve a P:Al ratio in the diet of 0.8:1 to 1.1:1. Cows consumed the RTM for at least 7 days to be eligible for blood collection. Blood samples were collected from the coccygeal vessel at the base of the tail and analyzed to determine blood phosphorus concentrations.

[0062] Based on a dietary P:Al ratio of 0.8:1 to 1.1:1, pre- and post-partum dairy cows had blood P concentrations in the range of 1.8 to 3 mg / dL. If blood P values ​​fell below 1.8 mg / dL, a new sample of the total mixed ration (TMR) was collected and the diet was confirmed again, as the dietary P:Al ratio was below 0.8:1 and needed to be reformulated. If blood P concentrations were above 3 mg / dL, a new TMR sample was collected to confirm the dietary P:Al ratio, as a P:Al ratio greater than 1.1:1 required reformulation of the diet to achieve the 0.8:1 to 1.1:1 P:Al ratio.

[0063] The methods defined above were conducted on 15 dairy farms that fed dairy cows pre-partum with zeolite A, and the data are shown in Fig. 2. Additional information on zeolite is presented in Table 1. Table 1 presents the responses of blood calcium (Ca) and phosphorus (P) levels to feeding dairy cows peri-partum with zeolite A, an aluminum-based product. Due to the diversity and multiplicity of data points, both pre- and post-partum, a single measurement of Ca and P in the blood, the closest to calving, is presented, with the responses to zeolite A feeding expressed as a percentage of the comparative treatment. lRutçth>87(E!60053ffiB, dte03 / (W>,2ffi!6, p. 6ΰ / &8 Clinical Milk Fever % of Control -33% 0% NR NR NR -75% ^po** OO τι NR -51% 0% NR Blood Phosphate Response % of Control NR -50% -47% -49% -73% >p O τι -72% -22% -45% sP 0s CD CO 1 0% Blood Calcium Response % of Control +27% +22% +11% +17% +13% +33% +57% +11% +13% +12% xp CO + Dietary Phosphate % DM Control 0.45 0.39 NR NR NR 69'0 69'0 0.38 0.43 0.3 00 o Dietary Phosphate % DM Zeolite 0.64 0.38 NR NR NR 0.61 0.61 0.42 0.36 0.3 00 o~ Dietary Ca % DM Control 0.45 CO CD o' NR NR NR 69'0 0.33 00 00 o' 2.53 CD o 2.79 Dietary Ca % DM Zeolite 0.64 0.65 NR NR NR 0.61 0.61 0.42 0.57 0.6 2.79 Treatments Zeolite vs Control Zeolite vs Control Zeolite vs DCAD Zeolite vs Control Zeolite vs Control Zeolite vs Control Zeolite vs Control Zeolite vs Control Zeolite vs Control Zeolite vs DCAD Zeolite vs Control Zeolite vs Control Reference Thilsing-Hansen et al., 2001 Kerwin et al., 2019 Frizzarini et al., 2022 Crookenden et al., 2020 Pallesen et al., 2007 Grabber et al., 2008 Saraiva de Oliveira, 2021 Thilsing-Hansèn et al, 2002 Khadouf et al., 2019 Λ / R = not reported. Petition 870260053893, dated 03 / 06 / 2026, pp. 51 / 66 Petition 870260067035, dated 07 / 07 / 2026, p. 62 / 118 19 / 19

[0064] The data demonstrate the effectiveness of the method, since blood phosphorus (P) levels are highly predictable based on the P:Al ratio provided. The data also corroborate feeding cows pre-partum with a dietary P:Al ratio of 0.8:1 to 1.1:1 to achieve peripartum blood P levels of 1.8 to 3.3 mg / dL. Conclusion:

[0065] Taken together, an aluminum-containing product, such as zeolite A, improves calcium homeostasis and blood calcium levels in periparturient cows by manipulating P homeostasis and inducing mild hypophosphatemia. The methodology presented here demonstrates a process for inducing transient hypophosphatemia in periparturient dairy cattle by manipulating the dietary P:Al ratio while providing an aluminum-containing product to achieve a dietary P:Al ratio of 0.8:1 to 1.1:1, which has been shown to result in desired blood P concentrations of 1.8 to 3 mg / dL in periparturient dairy cows.

[0066] Although the present invention has been described in considerable detail with reference to certain preferred embodiments, other embodiments are possible. The steps described for the present methods, for example, are not intended to be limiting nor to indicate that each step is necessarily essential to the method, but rather are merely exemplary steps. Therefore, the scope of the appended features should not be limited to the description of the preferred embodiments contained in this invention. All references cited herein are incorporated by reference in their entirety. lRutçth>87(E!6005383?!, de07SO7 / 2026,,pág;. 63118

Claims

1 / 7 CLAIMS 1. A method for reducing the risk of periparturient hypocalcemia in at least one ruminant animal during the periparturient period, characterized in that it comprises the steps of: a) measuring a phosphorus (P) concentration in the blood of at least one ruminant animal during the periparturient period; and b) administering orally to the ruminant animal during the periparturient period, during at least part of a pre-partum period of a ruminant animal during the periparturient period, a composition comprising: i) an effective amount of a phosphorus-binding compound to control the phosphorus concentration in the blood in a range of 1.5-3.5 mg P per dL of blood in the ruminant animal during the periparturient period, wherein the phosphorus-binding compound comprises an aluminum compound; wherein step a) may be performed before or after step b).

2. A method for assessing whether a ruminant in the periparturient period is susceptible to treatment to reduce the risk of periparturient hypocalcemia with a composition during at least part of the ruminant's prepartum period, the method being characterized in that it comprises measuring the concentration of phosphorus (P) in the blood in a sample from at least one ruminant in the periparturient period; wherein said composition comprises an aluminum compound in an amount to control the concentration of phosphorus in the blood in a range of 1.5 to 3.5 mg of P per dL of blood in the ruminant in the periparturient period.

3. Method, according to claim 1 or 2, characterized in that the concentration of phosphorus in the blood of lPeitçã(>87(E!60053ffiB, dte03 / (W>,2ffi!6,|piig. (5B118 2 / 7 step a) is controlled to be in a range of 1.8 to 3.0 mg of P per dL of blood; preferably a range of 2.0 to 3.0 mg of P per dL of blood.

4. Method according to claim 1 or 2, characterized in that: if the blood phosphorus concentration is below the range, the administration rate of the composition in step b) is reduced or the composition is reformulated to decrease the amount of the compound and / or increase the phosphorus to aluminum ratio; and, if the blood phosphorus concentration is above the range, the administration rate of the composition in step b) is increased or the composition is reformulated to increase the amount of the compound relative to the phosphorus in the composition and / or decrease the phosphorus to aluminum ratio, so that the blood phosphorus concentration is brought into the range.

5. Method according to claim 1 or 2, characterized in that the measurement of step a) is performed periodically; optionally in that the phosphorus-binding compound of step b) reduces the bioavailability of phosphorus in the digestive system of the ruminant animal during the periparturient period; optionally in that the composition of step b) is an animal feed composition, and the pharmacologically acceptable vehicle comprises one or more animal feed components other than the phosphorus-binding compound.

6. Method, according to claim 1 or 2, characterized in that the ruminant animal in the periparturient period is a dairy cow.

7. Method, according to claim 1 or 2, lRutçth>87(E!600538S5!, de07SO7 / 2026,,pg;. 65 / 118 3 / 7 characterized in that the compound further comprises a pharmacologically acceptable vehicle.

8. Method according to claim 7, characterized in that: a) the composition is a composition for animal feed; b) the pharmacologically acceptable carrier comprises one or more animal feed components other than the phosphorus binding compound; and c) the animal feed composition provides a weight ratio of phosphorus (P) to aluminum (Al) in the diet in a range of 0.7:1 to 1.4:

1.

9. Method according to claim 8, characterized in that the weight ratio is in a range of 0.8:1 to 1.2:

1.

10. A method according to claim 1 or 2, characterized in that the aluminum compound comprises an aluminosilicate, aluminum sulfate, aluminum chloride, aluminum ammonium sulfate, aluminum potassium sulfate, aluminum sodium sulfate, aluminum calcium silicate, aluminum hydroxide, aluminum bromide, aluminum iodide, aluminum nitrate, aluminum carbonate, a hydrate thereof, or any mixture thereof.

11. A method according to claim 1 or 2, characterized in that the aluminum compound comprises aluminum sulfate, aluminum chloride, aluminum hydroxide, a hydrate thereof, or any mixture thereof.

12. Composition for reducing the risk of peripartum hypocalcemia in a ruminant animal during the periparturient period, characterized in that it comprises: a) a phosphorus-binding compound present in an amount that controls the concentration of phosphorus in the blood of the ruminant animal during the periparturient period in a range of 1.5-3.5 mg of P lRutçã(>87(E!60053ÍSB, dte 07072026,,^. 66 / 118 4 / 7 per dL of blood, wherein the phosphorus-binding compound comprises an aluminum compound.

13. Use of a composition, as defined in claim 12, characterized in that it is for the manufacture of a feed composition, an oral solution, preferably an aqueous solution, or a dry formulation to reduce the risk of peripartum hypocalcemia in a ruminant animal during the peripartum period.

14. Use of: a) a phosphorus chelating compound, preferably wherein the phosphorus chelating compound comprises an aluminum compound; and optionally b) a pharmacologically acceptable vehicle and / or one or more feed components, characterized in that it is for the manufacture of a composition, a feed composition, an oral solution, preferably an aqueous solution, a dry formulation (e.g., a powder, a tablet, a capsule or similar) or a product to reduce the risk of periparturient hypocalcemia in a ruminant animal during the periparturient period.

15. Composition according to claim 12, or use according to claim 13 or 14, characterized in that the composition further comprises a pharmacologically acceptable vehicle.

16. Composition according to claim 12, or use according to claim 13 or 14, characterized in that the composition is an animal feed composition and the pharmacologically acceptable vehicle comprises one or more animal feed components other than the phosphorus binding compound.

17. Composition or use, according to claim 16, characterized in that the animal feed composition is suitable for a dairy cow.

18. Composition according to claim 12, or use according to claim 13 or 14, characterized in that the aluminum compound comprises an aluminosilicate, aluminum sulfate, aluminum chloride, aluminum ammonium sulfate, aluminum potassium sulfate, aluminum sodium sulfate, aluminum calcium silicate, aluminum hydroxide, aluminum bromide, aluminum iodide, aluminum nitrate, aluminum carbonate, a hydrate thereof, or any mixture thereof.

19. Composition, according to claim 12, or use, according to claim 13 or 14, characterized in that the animal feed composition provides a weight ratio of phosphorus (P) to aluminum (Al) in the diet in a range of 0.7:1 to 1.4:

1.

20. Method for reducing the risk of peripartum hypocalcemia in at least one ruminant animal during the periparturient period, characterized in that it comprises the steps of: a) measuring the aluminum concentration in an animal feed composition, wherein the animal feed composition comprises: i) an effective amount of an aluminum compound to control the blood phosphorus concentration in a range of 1.5 to 3.5 mg P per dL of blood in the ruminant animal during the periparturient period;b) before or after step a), administer orally to the ruminant animal during the periparturient period, for at least part of the pre-partum period of the ruminant animal in the periparturient period, the animal feed composition, wherein the animal feed composition comprises: c) if the aluminum concentration is below a range of 0.25 to 0.40% (or 2500 to 4000 ppm), the administration rate of the feed composition from step b) is decreased or the animal feed composition is reformulated to decrease the amount of aluminum compound and / or increase the phosphorus to aluminum ratio;(e, d) if the blood phosphorus concentration is above the range, the feeding rate of the animal feed composition from step b) is increased or the animal feed composition is reformulated to increase the amount of aluminum compound relative to phosphorus in the composition and / or decrease the phosphorus-to-aluminum ratio, so that the blood phosphorus concentration is brought into the range.

21. Method for assessing whether a ruminant in the periparturient period is susceptible to treatment to reduce the risk of periparturient hypocalcemia by controlling the amount of feed intake during at least part of the prepartum period, the method being characterized in that it comprises the steps of: a) measuring the concentration of aluminum in a feed composition, wherein the feed composition comprises: i) an effective amount of an aluminum compound to control the concentration of phosphorus in the blood in a range of 1.5 to 3.5 mg P per dL of blood in the ruminant in the periparturient period; b) if the aluminum concentration is below a range of 0.25 to 0.40% (or 2500 to 4000 ppm), the said treatment comprises decreasing the intake of the feed composition or reformulating the feed composition to decrease the amount of the aluminum compound and / or increase the phosphorus / aluminum ratio;and, c) if the blood phosphorus concentration is above the range, the aforementioned treatment comprises increasing the intake of the feed composition from step b) or reformulating the feed composition to increase the amount of aluminum compound relative to phosphorus in the composition and / or decrease the phosphorus / aluminum ratio, lPeitçth>87(E!600538SB, dte03 / (W>,2ffi!6, pág. 69 / 118 7 / 7 so that the blood phosphorus concentration is brought into the range.; 22. Method according to claim 20 or 21, characterized in that the concentration of phosphorus in the blood in step b) is controlled to be in a range of 1.8-3.0 mg P per dL of blood; preferably in a range of 2.0-3.0 mg P per dL of blood.

23. Method according to claim 22, characterized in that the measurement of step a) is performed periodically; optionally wherein the animal feed composition further comprises a pharmacologically acceptable carrier which is one or more components of the animal feed other than the phosphorus binding compound; optionally wherein the ruminant animal in the periparturient period is a dairy cow. lRutçth>87(E!600538SB, dteST / lTíflBEa^.gBg;. 70 / 118

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