Improvement of apparatus and methods for delivering substances to animals

Abstract

The present invention provides delayed-release dosage forms and boluses configured for administration to animals. It also provides the use of the delayed-release dosage forms or boluses of the present invention to reduce methane production in ruminants. Methods for producing the boluses of the present invention are also provided. [Solution] A delayed-release dosage form or bolus configured for administration to an animal, wherein the dosage form and the bolus are configured to release a hydrophobic substance to the animal over a period of time.

Description

Technical Field

[0001] The present invention relates to improvements in devices and methods for animal production and the delivery of substances to animals, and more particularly to devices and methods for administering at least one beneficial substance to animals, and a method of manufacturing such devices.

Background Art

[0002] In agriculture, it is often necessary to deliver substances to animals. This has various purposes, including but not limited to treating and preventing diseases and increasing animal production.

[0003] There are various devices and methods for delivering substances such as drugs to animals. However, one type of compound that is difficult to deliver to animals is hydrophobic compounds. The properties of these compounds pose challenges, particularly in the development of controlled release technologies for these hydrophobic substances through the animal's stomach.

[0004] One specific objective of administering substances to animals is to reduce the adverse effects of agriculture. For example, various methane and nitrification inhibitors are known to be administered to animals to reduce or mitigate the adverse effects of methane and nitrate-containing compounds produced by animals.

[0005] However, despite current efforts, climate change is having a widespread impact on the environment and society worldwide. It is widely understood that these impacts continue to increase over time. As a result, globally, there is a push to reduce harmful greenhouse gas (GHG) emissions in order to avoid the worst effects of climate change.

[0006] The agricultural sector is considered a major source of GHG emissions. The total methane emissions from livestock worldwide are estimated to be 7.1 gigatons per year in terms of CO2 equivalent, corresponding to 14.5% of total anthropogenic GHG emissions. Therefore, this sector will play an important role in reducing overall GHG emissions.

[0007] The main GHGs released by agriculture are methane (CH4) and nitrous oxide (N2O), with livestock being the primary source of methane emissions. Most of the methane is released when cattle burp. The amount of methane produced on each farm is directly related to the total amount of feed consumed by the animals.

[0008] Countries with strong agricultural sectors, such as New Zealand, face the challenging goal of reducing agricultural emissions. For example, the New Zealand government has introduced policies aimed at reducing methane emissions by 24-50% by 2050. In New Zealand, livestock methane production is estimated to account for half of the country's total greenhouse gas emissions. Reducing methane is a crucial element in achieving greenhouse gas emission targets and mitigating the effects of global warming.

[0009] GHG emissions by animals also negatively impact animal productivity. Feed that is converted into compounds and then expires or is released by animals is an energy source that has not been converted into a productive use. Therefore, to increase efficiency, it is important to optimize the conversion of feed into animal productivity in the form of weight gain and milk production. [Overview of the project] [Problems that the invention aims to solve]

[0010] The object of the present invention is to provide improved apparatus and methods for delivering substances to animals, such as hydrophobic substances and / or methane inhibitors.

[0011] The object of the present invention is to provide an apparatus and method for reducing GHG emissions.

[0012] The object of the present invention is to provide an apparatus and method for improving or optimizing the productivity of animals.

[0013] Alternatively, an object of the present invention is to provide apparatus and methods for improving the productivity of animals, for example, by reducing methane production.

[0014] The object of the present invention is to provide a formulation for reducing GHG emissions by one or more animals, such as ruminants.

[0015] The object of the present invention is to provide an apparatus and method that can release a substance at different rates over a certain period of time.

[0016] Alternatively, an object of the present invention is to provide a method for manufacturing a device for delivering substances, such as substances that reduce the emission of GHGs, to animals.

[0017] Alternatively, an object of the present invention is to overcome some of the shortcomings of the prior art.

[0018] Alternatively, an object of the present invention is to provide the public with a useful alternative. [Means for solving the problem]

[0019] According to one aspect of the present invention, a bolus configured for administration to an animal is provided, which is configured to release a hydrophobic substance to the animal over a period of time.

[0020] According to one aspect of the present invention, a bolus for administration to a ruminant is provided, wherein the bolus is configured to release an effective amount of a substance, which is preferably at least one inhibitor.

[0021] A further aspect of the present invention provides a method for reducing gas emissions (preferably methane) from ruminants, the method comprising the step of administering a bolus containing at least one inhibitor to the ruminant.

[0022] According to another aspect of the present invention, there is provided the use of a methane inhibitor and a carrier in a bolus for reducing methane production in ruminants.

[0023] According to another aspect of the present invention, there is provided the use of a methane inhibitor and a carrier in a bolus for reducing methane emissions from ruminants.

[0024] According to another aspect of the present invention, there is provided the use of haloform in the manufacture of a bolus for reducing the emission of one or more greenhouse gases ("GHGs") from ruminants.

[0025] In a preferred embodiment, the bolus may be configured to be administered to a ruminant, and the ruminant may include beef or dairy cattle, sheep, goats, buffalo, deer, elk, giraffe or camel.

[0026] In one embodiment, the bolus may be adapted to reduce the release of one or more greenhouse gases ("GHGs") from ruminants.

[0027] In another embodiment, the bolus may be a sustained-release bolus configured to release at least one inhibitor over a period of time, for example, within the rumen of the animal.

[0028] According to a further aspect, there is provided a bolus for administration to a ruminant, the bolus comprising: a core comprising at least one substance for administration to a ruminant mixed with a carrier; and a housing covering at least a portion of the core, the bolus being configured to release the substance through the housing over a period of time.

[0029] In another aspect of the present invention, there is provided: a core containing a substance for administration to an animal, and a bolus comprising a housing at least partially covering a portion of the core, The housing is formed from at least one polylactic acid (PLA) material.

[0030] In a further aspect of the present invention, a bolus is provided which includes a core, the core comprising a mixture of at least one wax and haloform.

[0031] The inventors have surprisingly discovered that the technology described herein may offer numerous advantages. These advantages may be the result of unique synergistic interactions between various aspects of the technology. Therefore, the technology of the present invention is described based on the inventors' current understanding of these interactions. It should be understood that any aspect, or two or more aspects, of the interaction described herein may form a separate invention.

[0032] Throughout this specification, the term “substance” or “substance administered to an animal” should be understood to mean any substance that benefits an animal, such as a drug for the treatment or prevention of disease, thereby improving animal productivity and mitigating at least one adverse effect on agriculture.

[0033] In a preferred embodiment, the substance may be a hydrophobic substance.

[0034] In particularly preferred embodiments, the hydrophobic substance may be an inhibitor. This specification refers to substances as inhibitors. However, this should not be considered limiting the scope of the invention, and alternatives are conceivable, such as hydrophilic substances.

[0035] In one embodiment, at least one inhibitor may be a methane inhibitor. The use of methane inhibitors can offer several advantages. For example, methane inhibitors reduce or eliminate methane production by ruminants, for instance, in the rumen. As a result, the amount of methane in the rumen that could be released by ruminants is reduced, and therefore GHG emissions are effectively reduced.

[0036] Furthermore, reducing methane production can potentially benefit animal production. For example, reducing methane means that a relatively large portion of the ingested feed is digested and converted into protein (milk or meat). As a result, farmers may be able to achieve higher productivity relative to the amount of feed given, or improve efficiency by reducing feed accordingly.

[0037] In one embodiment, the methane inhibitor may be a haloform.

[0038] In a preferred embodiment, the methane inhibitor may be selected from the list of chloroform, bromoform, iodoform, or a combination thereof.

[0039] In a particularly preferred form, the haloform may be bromoform (CHBr3). Using bromoform can offer several advantages. For example, it can be highly effective at relatively low doses, and a sufficient amount of the inhibitor can be delivered over a long period of time with a single device. Furthermore, because bromoform is relatively dense, the total weight of the bolus increases, causing it to remain within the lumen, i.e., sink into the ventral part of the lumen rather than floating, thus reducing backflow.

[0040] However, despite these advantages, the inventors have faced numerous challenges and problems in developing boluses for the controlled release of haloforms, particularly bromoform, to ruminants.

[0041] In further embodiments, the bolus may include a core.

[0042] The core can be formed by an inhibitor mixed with a carrier.

[0043] However, in another embodiment, the inhibitor may be provided in a substantially pure form, for example, a purified form, and not mixed with a carrier, for example.

[0044] In embodiments, the support may have a structure that promotes or enhances the affinity of the inhibitor to the support. For example, the support may have polar functional groups.

[0045] In embodiments, the support may be a relatively polar substance, for example, having a relatively high %w / w polar functional group. Surprisingly, the inventors discovered that the support and the inhibitor can interact, and that this interaction can affect the release rate of the inhibitor from the bolus. This aspect of the present invention should become clearer from the following description.

[0046] Examples of suitable functional groups to be included in the support are esters, fatty acids, aliphatic alcohols, carbonyls, and aliphatic amines. While not limited to a specific mechanism, the inventors believe that inhibitors may interact with polar functional groups in the wax potentially through the formation of hydrogen bonds. The amount of polar functional groups present in the support influences the affinity between the support and the inhibitor.

[0047] The inventors have discovered that a certain range of substances may be suitable for use as a carrier in the present invention. For example, the carrier may be selected from the list of waxes, myristic acid, stearic acid, steryl alcohols, cetyl alcohols, cetosteryl alcohols, or combinations thereof.

[0048] In a particularly preferred embodiment, the carrier may be a waxy substance. For example, the carrier may be selected from the list of beeswax, paraffin wax, PEG4000, carnauba wax, castor wax, candelilla wax, jojoba wax, or lanolin, or a combination thereof.

[0049] In particularly preferred embodiments, the carrier may include paraffin wax and castor wax.

[0050] In a particularly preferred embodiment, the carrier may contain paraffin wax and castor wax in a ratio of about 50:50 (parts by weight).

[0051] In another embodiment, the support may comprise a mixture of two or more components. For example, the support may comprise a mixture of at least one relatively polar substance and a relatively nonpolar substance. For example, in some forms, the support may comprise a mixture of paraffin wax (a mixture of alkanes without polar functional groups) and castor wax and / or carnauba wax (which has a relatively large amount of polar functional groups). As a result, the overall polarity of the support can be adjusted to achieve a desired affinity for the inhibitor. This can then be used to achieve a desired release rate of the inhibitor.

[0052] Furthermore, in addition to the above, solid carriers such as powdered activated carbon, zeolite, and bentonite can also be used as carriers. Therefore, the discussion herein should not be considered to limit the scope of the present invention.

[0053] In further embodiments, the carrier may include one or more additional components. For example, additional components such as zinc or zinc oxide may be incorporated. Preferably, a high-density material such as a metal piece (preferably steel) may be included in the carrier. The additional components may be used to achieve a desired density of the core and / or bolus.

[0054] It should also be understood that additional components may be added to the bolus cavity separately from the support, without mixing with the support. This is particularly beneficial for forming a core with a desired release profile, and the density of the bolus can be adjusted to a desired amount by including additional components.

[0055] Other suitable additives for incorporation into the carrier may include colloidal silicon dioxide, charcoal, bentonite, and zeolite.

[0056] Further embodiments of the carrier, and their effect on the release of inhibitors from the bolus, along with the interaction between the carrier and the housing, should become clearer from the following description.

[0057] In preferred embodiments, the carrier may have a melting point substantially between 50 and 90°C.

[0058] In a particularly preferred embodiment, the support has a melting point lower than the boiling point of the inhibitor. This can be useful because it allows the support to be melted and mixed with the inhibitor without substantial loss of the inhibitor due to evaporation.

[0059] In a preferred embodiment, the core may have a melting point higher than 37°C.

[0060] In a particularly preferred embodiment, the core may have a melting point above 40°C.

[0061] The melting point of the core may be beneficial to the functionality of this technology in several ways. For example, having a melting point of 37°C, more preferably above 40°C, can help the carrier stabilize the inhibitor when the bolus is in the lumen. This may be beneficial for controlling the release of the inhibitor, for example, the migration of the inhibitor through the material forming the housing.

[0062] In one embodiment, the bolus may be adapted to achieve a maximum release rate of approximately 0.05 g to 2 g of bromoform per day into the lumen.

[0063] In one embodiment, the bolus may be adapted to release between 0.02 g and 0.5 g of bromoform into the lumen per day.

[0064] In a particularly preferred embodiment, the bolus may be adapted to achieve a maximum release rate of approximately 0.1 to 0.5 g of bromoform per day into the lumen.

[0065] In a preferred embodiment, the bolus is configured to release 0.02 g to 0.3 g of bromoform into the lumen per day.

[0066] In one embodiment of the present invention, the core of the bolus may contain haloform, preferably bromoform, in an amount of 30% to 80% by weight, preferably 55% to 75% by weight, and more preferably 50% by weight.

[0067] In a particularly preferred embodiment, the core contains a haloform, preferably bromoform, at a concentration of 55% (by weight) or less.

[0068] The inventors have discovered that the rate of release of the inhibitor into the lumen increases over time. This may be the result of several factors. Thus, the release rate starts at zero upon administration to animals and increases to a maximum. However, the above should not be considered limiting, and other release rates are also considered to be within the scope of the invention.

[0069] In further embodiments, the bolus may include a housing.

[0070] Throughout this specification, references to the term “housing” should be understood to mean a structure capable of receiving and holding a core containing at least one inhibitor.

[0071] In a preferred embodiment, the housing comprises a body having a cavity in which the core is located.

[0072] However, it should be understood that the housing may take on other shapes. For example, the housing may include two or more cavities, each capable of receiving and holding a separate core.

[0073] In one embodiment, the housing may include an open end.

[0074] The bolus can be used with an open end, for example, by administering it to an animal with the end open. As a result, in these embodiments, the open end provides an opening for exposing the contents of the core to the fluid in the lumen during use.

[0075] In yet another preferred embodiment, the housing can completely enclose and surround the core, for example having a sealed cavity in which the core is placed.

[0076] For example, a bolus may include a housing comprising a cavity in which at least a portion of the core may be placed, and an open end to facilitate insertion of the core into the cavity. The open end can be covered with a cap.

[0077] The cap may be formed separately from the housing and may be removable or permanently fixed to the housing. Alternatively, the cap may be formed integrally with the housing.

[0078] In yet another embodiment, the housing may consist of at least two parts, each having a cavity for receiving a corresponding part of the core. Together, the at least two parts completely enclose the core, defining a sealed cavity in which the core is placed.

[0079] In yet another embodiment, the housing may be formed around the core, for example, by molding. Alternatively, the housing and cap together may define a substantially closed and sealed cavity in which the core is placed.

[0080] The inventors believe it is preferable to provide a substantially or completely closed and sealed cavity, as this can help achieve the desired controlled release of the inhibitor from the bolus of the present invention. For example, in such embodiments, the inhibitor can pass through the material forming the housing, for example, by mass diffusion.

[0081] In some embodiments, the housing may be configured to have sufficient structural integrity to remain undamaged for a predetermined period of time.

[0082] In a preferred embodiment, the housing may be configured to be disassembled over a predetermined period of time.

[0083] Throughout this specification, any reference to the term “a specified period” should be understood to mean the period during which the inhibitor is released to the animal.

[0084] In a particularly preferred embodiment, the predetermined period may be at least 2 months, preferably 6 months, and more preferably 12 months.

[0085] The inventors have surprisingly discovered that the housing of the present invention may facilitate the controlled release of the inhibitor. For example, the housing can withstand the conditions in the lumen for a predetermined period. During this time, the housing protects the core from the liquid in the lumen, but can also facilitate or control the controlled release of the inhibitor. However, due to the design of the housing, the housing may disintegrate or decompose over a predetermined period. This can contribute to mitigating adverse effects from device administration to animals and ensuring that animals are treated with multiple boluses, for example, a second bolus may be administered at the end of a predetermined period, towards the end, or after the end.

[0086] In embodiments of the present invention, the thickness of the housing can be selected to contribute to the release rate of the inhibitor. For example, the inventors have found that the thickness of the housing can affect the release rate of the inhibitor from the bolus. In these embodiments, a relatively thick housing results in a relatively slower release rate than a relatively thin housing.

[0087] In a preferred embodiment, the housing may have a thickness of at least 1 mm.

[0088] In yet another preferred embodiment, the housing may have a thickness of less than 3 mm.

[0089] In yet another preferred embodiment, the housing may have a thickness between 1.5 and 2 mm, or between 0.5 and 2 mm.

[0090] In a particularly preferred embodiment, the housing has a thickness of 1 mm.

[0091] The thickness of the housing can be particularly important in embodiments such as those in which the core is completely enclosed by the housing, in order to achieve the desired controlled release of the inhibitor. This should become clear from the following discussion.

[0092] In one embodiment, the dimensions of the cavity may vary along the length of the housing.

[0093] In a preferred embodiment, the cavity includes at least two regions having different cross-sectional areas, for example, a first region having a first cross-sectional area and a second region having a second cross-sectional area.

[0094] In a particularly preferred embodiment, the first region has a relatively small cross-sectional area, and the second region has a relatively large cross-sectional area.

[0095] In yet another preferred embodiment, the first region may be located closer to the open end than the second region.

[0096] Having cavities with regions having different cross-sectional areas facilitates more controlled release of the inhibitor, better meeting the animal's requirements. For example, a relatively small cross-sectional area can be provided near the open end to deliver a relatively small dose of the inhibitor, while a relatively large cross-sectional area may be provided near the distal end, which can be useful, for example, when the dosage of the inhibitor needs to be increased over time due to the animal's growth.

[0097] It should be understood that the opposite configuration may also be provided; for example, a relatively large cross-sectional area may be located near the open end, and a relatively small cross-sectional area near the distal end. This configuration may be useful when a higher dose of the inhibitor is desired initially, followed by lower doses. For example, this configuration can be used when the animal has a high demand for the inhibitor, such as during periods of relatively high feed intake and energy requirements, such as during milking, or during dry periods followed by periods of relatively low feed intake.

[0098] Furthermore, it should be understood that the cross-sectional area of ​​the cavity can increase gradually and continuously from the first region to the second region; for example, there is no defined "step" between the first and second regions.

[0099] In other embodiments, the housing may include a third region having a third cross-sectional area. This may be further used to control the dosage of the inhibitor to an animal. Therefore, the foregoing should not be considered to limit the scope of the art.

[0100] In one embodiment, the thickness of the housing walls may vary along the length of the housing. In such an embodiment, the thickness of the wall at or toward one end of the housing may be greater than the thickness of the wall at the distal end. For example, the thickness of the wall at or toward the open end may be less than that of the distal end.

[0101] This arrangement can be particularly beneficial for controlling the release of inhibitors over time. For example, a relatively thin wall decomposes relatively faster than a relatively thick wall. This structure can be used to control the rate of decomposition along the length of the housing. For example, it can be used to ensure that the open end is the only place where the liquid in the lumen can come into contact with and erode the core.

[0102] In a preferred embodiment, the housing is made from a material that allows the inhibitor to migrate during use, for example, by a mass diffusion process.

[0103] In a preferred embodiment, the housing may be made from at least one plastic material. For example, the housing may be made from a biodegradable plastic or material that decomposes over time within the lumen.

[0104] In particularly preferred embodiments, the housing may be made from a material selected from one or more of the following: polylactic acid (PLA), polyglycolic acid (PGA), polyglycolic acid polylactic acid (PLGA), polypropylene, polycaprolactone (PCL), poly(d-lactic acid) (PDLA), polybutylene succinate (PBS), polybutylene adipate terephthalate (PBAT), SLA polymer, ABS, or combinations thereof. In particularly preferred embodiments, the housing comprises PLA and PBS.

[0105] The housing material may contain PLA, PBAT, and / or PBS in different proportions (by weight %) as shown in Examples 1-7 of the table below. [Table 1]

[0106] In a particularly preferred embodiment, the housing material comprises PLA and PBS in a weight ratio ranging from 100:0 to 40:60 PLA:PBS.

[0107] In a particularly preferred embodiment, the housing comprises PLA and PBS in a weight ratio ranging from 100:0 to 40:60 PLA:PBS, and the housing has a thickness between 0.4 and 1.5 mm.

[0108] In further embodiments, the core of the bolus of the present invention is covered by a plurality of housings arranged concentrically (for example, like an onion). Such a plurality of housings (e.g., two, three, or more housings) has the advantage that the bolus does not decompose (e.g., by wear) in the lumen as quickly. As a result, the haloform in the core persists longer in the lumen, and methane production is reduced over a longer period. In embodiments with a plurality of housings, the material and thickness of the housings may be as described herein for other embodiments. In preferred embodiments, the bolus of the present invention comprises at least two housing layers, one outer housing and one inner housing, wherein the material of each housing comprises a biodegradable polymer, preferably a biodegradable polymer selected from the group consisting of polylactic acid (PLA), polybutylene succinate (PBS), polybutylene adipate terephthalate (PBAT), and combinations thereof.

[0109] Furthermore, the housing may be made of non-biodegradable materials such as EVA, silicon, or acrylate. Consequently, the discussion herein should not be considered to limit the scope of the present invention.

[0110] Furthermore, the housing material may contain one or more other compounds, such as plasticizers, hardeners, and colorants.

[0111] However, in alternative embodiments, the housing may be made of one or more non-adsorbent materials, i.e., materials from which the inhibitor does not migrate into or through. Using non-absorbent materials for the housing can help control the release rate of the inhibitor in certain embodiments, such as open-end boluses. For example, in these embodiments, the concentration of the inhibitor in the core is not reduced by absorption into the housing material.

[0112] In some embodiments, the bolus may include a barrier layer. In these embodiments, the barrier layer may be placed between at least a portion of the core and the housing. For example, the barrier layer can minimize or completely prevent contact between the portion of the core and the housing. This can help prevent the dissolution of the inhibitor (or other compounds), thereby better controlling the release of the inhibitor and improving the stability of the device. This may be particularly useful when the solubility of the inhibitor is high in the material from which the housing is made.

[0113] Alternatively, in embodiments where the barrier layer is provided only between a portion of the core and the housing, the movement of the inhibitor into the housing can be reduced, but not completely prevented. In fact, because the barrier layer reduces the contact area between the core and the housing, it may reduce the rate of inhibitor release compared to when the barrier layer is not provided.

[0114] Alternatively, the bolus may not include a barrier layer. This configuration may be useful when the solubility of the inhibitor in the materials constituting the housing is relatively low. It may also be useful when the composition of the housing and / or carrier is selected to control the release rate, for example, the diffusion rate of the inhibitor through the housing.

[0115] In another embodiment, the bolus may be adapted to have a core and housing dissolution rate that results in substantially uniform dissolution of both components over time within the lumen.

[0116] In one embodiment, a cavity within the housing may provide a reservoir configured to accept a certain amount of inhibitor. For example, the reservoir may be a closed cavity within the housing that can accept and hold a certain amount of inhibitor.

[0117] In one embodiment, the bolus may include a dispensing mechanism.

[0118] In one embodiment, the carrier may have a relatively high affinity for the inhibitor compared to the affinity of the housing for the inhibitor. As discussed elsewhere in this document, this can be achieved by the relative polarity of the materials forming the carrier and housing, and by appropriately matching these materials to the inhibitor.

[0119] In another embodiment, the housing may be formed from a material having a Shore D hardness of at least 40. In such embodiments, it is believed that having a housing with a Shore D hardness lower than 40 would result in a bolus that is too soft, which could hinder bolus delivery to animals or cause damage or premature degradation of the animals before the full amount of the inhibitor is administered.

[0120] In further embodiments, the housing may be formed from a material having a Shore D hardness of less than 80.

[0121] In another embodiment, the housing may be configured to facilitate controlled release of the inhibitor from the core. While not limited to a specific mechanism, the inventors hypothesize that the inhibitor may be released through the housing by a mechanism of mass diffusion.

[0122] In a further embodiment, the present invention provides a bolus for administration to a ruminant, the bolus comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a housing covering at least a portion of the core, or preferably the entire core, wherein the bolus is configured to release the haloform.

[0123] In a further embodiment, the present invention provides a bolus for administration to ruminants, the bolus comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier, and a housing covering at least a portion or all of the core, wherein the bolus is configured to release the haloform; the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof.

[0124] In a further embodiment, the present invention provides a bolus for administration to ruminants, the bolus comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a housing covering at least a portion of the core, preferably the entire core; wherein the bolus is configured to release the haloform, the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; and the housing comprising a biodegradable polymer, preferably a biodegradable polymer selected from the group consisting of polylactic acid (PLA), polybutylene succinate (PBS), polybutylene adipate terephthalate (PBAT) and combinations thereof.

[0125] In a further embodiment, the present invention provides a bolus for administration to ruminants, the bolus comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a housing covering at least a portion of the core, preferably the entire core; wherein the bolus is configured to release the haloform; and the carrier comprises a wax, preferably castor wax, paraffin wax, or a mixture thereof; and the housing has a layer thickness between 0.4 and 1.5 mm.

[0126] In a further embodiment, the present invention provides a bolus for administration to ruminants, the bolus comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a housing covering at least a portion of the core, or preferably the entire core; wherein the bolus is configured to release the haloform; and the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; and the housing comprising a biodegradable polymer, preferably a biodegradable polymer selected from the group consisting of polylactic acid (PLA), polybutylene succinate (PBS), polybutylene adipate terephthalate (PBAT) and combinations thereof; and the housing having a layer thickness between 0.4 and 1.5 mm.

[0127] In a further embodiment, the present invention provides a bolus for administration to ruminants, the bolus comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a housing covering at least a portion of the core, or preferably the entire core; the bolus being configured to release the haloform; the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; the housing comprising a biodegradable polymer, preferably a biodegradable polymer selected from the group consisting of polylactic acid (PLA), polybutylene succinate (PBS), polybutylene adipate terephthalate (PBAT), and combinations thereof; and the thickness of the housing layer being less than 2 mm.

[0128] In a further embodiment, the present invention provides a bolus for administration to ruminants, the bolus comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a housing covering at least a portion or all of the core; wherein the bolus is configured to release the haloform; the core further comprising at least one metal piece (such as a metal pellet and / or metal rod), the metal preferably being steel or zinc. An advantage of this embodiment is that the bolus density is increased and the likelihood of the bolus being backflowed by the animal is reduced. Preferably, the bolus of the present invention further comprises a densifier, preferably the densifier comprising at least one metal piece, preferably the densifier being provided within the core.

[0129] In a further embodiment, the present invention provides a bolus for administration to ruminants, the bolus comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a housing covering at least a portion of the core, preferably the entire core; wherein the bolus is configured to release the haloform; the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; the housing comprising polylactic acid (PLA); and preferably the thickness of the housing layer being less than 2 mm. In a further embodiment, the present invention provides a delayed-release dosage form for administration to ruminants, the delayed-release dosage form comprising a core comprising a haloform (preferably bromoform); and a coating covering at least a portion of the core, preferably the entire core, wherein the delayed-release dosage form is configured to release the haloform.

[0130] In a further embodiment, the present invention provides a delayed-release dosage form for administration to ruminants, the delayed-release dosage form comprising a core comprising a haloform (preferably bromoform); and a coating covering at least a portion or preferably the entire core; wherein the delayed-release dosage form is configured to release the haloform; the core further comprises a wax, preferably castor wax, paraffin wax, or a mixture thereof.

[0131] In a further embodiment, the present invention provides a delayed-release dosage form for administration to ruminants, the delayed-release dosage form comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a coating covering at least a portion of the core, or preferably the entire core; the delayed-release dosage form being configured to release the haloform; the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; and the coating comprising a biodegradable polymer, preferably a biodegradable polymer selected from the group consisting of polylactic acid (PLA), polybutylene succinate (PBS), polybutylene adipate terephthalate (PBAT), and combinations thereof.

[0132] In a further embodiment, the present invention provides a delayed-release dosage form for administration to ruminants, the delayed-release dosage form comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a coating covering at least a portion of the core, or preferably the entire core; the delayed-release dosage form being configured to release the haloform; and the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; and the coating having a layer thickness between 0.4 and 1.5 mm.

[0133] In a further embodiment, the present invention provides a delayed-release dosage form for administration to ruminants, the delayed-release dosage form comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a coating covering at least a portion of the core, or preferably the entire core; wherein the delayed-release dosage form is configured to release the haloform; the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; the coating comprising a biodegradable polymer, preferably a biodegradable polymer selected from the group consisting of polylactic acid (PLA), polybutylene succinate (PBS), polybutylene adipate terephthalate (PBAT), and combinations thereof; and the coating having a layer thickness between 0.4 and 1.5 mm.

[0134] In a further embodiment, the present invention provides a delayed-release dosage form for administration to ruminants, the delayed-release dosage form comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a coating covering at least a portion of the core, or preferably the entire core; wherein the delayed-release dosage form is configured to release the haloform; the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; the coating comprising a biodegradable polymer, preferably a biodegradable polymer selected from the group consisting of polylactic acid (PLA), polybutylene succinate (PBS), polybutylene adipate terephthalate (PBAT), and combinations thereof; and the thickness of the coating layer being less than 2 mm.

[0135] In a further embodiment, the present invention provides a delayed-release dosage form for administration to ruminants, the delayed-release dosage form comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a coating covering the core; wherein the delayed-release dosage form is configured to release the haloform; and the core further comprising at least one metal piece (such as a metal pellet and / or a metal rod), the metal preferably being steel or zinc. An advantage of this embodiment is the increased density of the delayed-release dosage form, which reduces the likelihood of the delayed-release dosage form being regurgitated by the animal.

[0136] In a further embodiment, the present invention provides a delayed-release dosage form for administration to ruminants, the delayed-release dosage form comprising: a core comprising a haloform (preferably bromoform) mixed with a carrier; and a coating covering part or all of the core; wherein the delayed-release dosage form is configured to release the haloform; the carrier comprising a wax, preferably castor wax, paraffin wax, or a mixture thereof; the coating comprising polylactic acid (PLA); and preferably the thickness of the coating layer being less than 2 mm. Experiments have shown that the thickness of the coating layer being less than 2 mm is preferable because this thickness allows the haloform to penetrate out of the core material at an optimal rate.

[0137] In the delayed-release formulation or bolus of the present invention, preferably less than 50% of the haloform contained in the core is released over a period of 3 months. In a preferred embodiment of the delayed-release formulation or bolus of the present invention, the core contains at least 100 grams of haloform. The core of the bolus or delayed-release formulation of the present invention preferably contains 30% to 70% by weight of haloform (preferably bromoform).

[0138] At present, it is understood that the controlled release of inhibitors through a housing can be affected by many factors. For example, the affinity of the inhibitor to the carrier can affect the diffusion of the inhibitor through the housing. It is understood that carriers with higher polarity or those containing a high degree of polar functional groups have a higher affinity for the inhibitor than carriers with lower polarity or those containing fewer functional groups.

[0139] The relative affinity of the materials forming the housing and core to the inhibitor may also affect the controlled release of the inhibitor from the core. For example, having a housing with a relatively low affinity for the inhibitor compared to the affinity of the carrier to the inhibitor can be a factor in controlling the rate of inhibitor release from the core. These aspects of the present invention should become clearer from the description herein.

[0140] Throughout this specification, references to the term “release mechanism” should be understood to mean a configuration for releasing a predetermined amount of inhibitor over time. For example, a release mechanism may comprise a valve device capable of releasing a certain amount of inhibitor through an outlet. Alternatively, a release mechanism may be a syringe-type mechanism comprising a plunger and an actuator; over time, the actuator moves the plunger in a reservoir, displacing the inhibitor from the reservoir.

[0141] Furthermore, the following items are also in accordance with the present invention. Item 1 provides a bolus for administration to ruminants, the bolus being configured to release an effective amount of at least one inhibitor. Item 2 provides any one bolus of Item 1, wherein at least one of the inhibitors is a methane inhibitor. Item 3 provides a bolus of item 1 or 2, and at least one inhibitor is a haloform selected from chloroform, bromoform, iodoform, or a combination thereof. Item 4 concerns one bolus from any of items 1-3, where at least one inhibitor is bromoform. Item 5, with respect to one of the boluses from Items 1-4, states that the bolus contains a core containing a certain amount of the inhibitor. Item 6 provides the bolus of Item 5, and the core contains a carrier mixed with the inhibitor. Item 7 relates to the bolus described in Item 6, wherein the carrier is a waxy substance selected from beeswax, paraffin wax, PEG4000, carnauba, candelilla, jojoba, or lanolin, or a combination thereof. Item 8 relates to one of the boluses from items 5-7, where the core has a melting point above 37°C. Item 9, relating to any one of items 5-8, states that the bolus includes a housing for receiving and holding the core. Regarding the bolus of item 9, item 10 states that the housing includes a cavity in which the core can be received and held. Item 11 relates to the bolus of item 9 or 10, and the housing includes an opening to facilitate exposure of the core to the fluid in the lumen of the ruminant animal during use. Item 12 relates to a bolus of item 10 or 11, and the cavity comprises a first region having a first cross-sectional area and a second region having a second cross-sectional area, wherein the first and second cross-sectional areas are different from each other to facilitate controlled release of the inhibitor from the core. Item 13 relates to one of the boluses described in items 9-12, wherein the housing is configured to be disassembled over a specified period of time. Item 14 relates to one of the boluses in items 9-13, and the housing is made from one or more non-adsorbent materials selected from: polylactic acid (PLA), polyglycolic acid (PGA), polyglycolic acid polylactic acid (PLGA), polypropylene, SLA polymer, PBS, or a combination thereof. Item 15 relates to any one of the boluses in items 9-14, further comprising a barrier layer between at least a portion of the housing and the core to isolate the portion of the housing from contact with the core. Item 16 relates to one of the boluses from items 1 through 15, which are adapted to release an inhibitor dose of approximately 0.1 g to 0.5 g per day into the rumen of a ruminant. Item 17 relates to one of the boluses from items 1 through 16, and the bolus is designed to release the inhibitor over a period of at least six months. Item 18 relates to one of the boluses from items 1 through 17, and the bolus is designed to release the inhibitor within two years. Item 19 provides a method for reducing gas emissions from ruminants, the method comprising the step of administering a bolus of any one of items 1 to 18 to the ruminant. Item 20 provides a method for reducing methane production in ruminants, the method comprising the step of administering a bolus of any one of items 1 to 18 to the ruminant. Item 21 provides the use of methane inhibitors and carriers in boluses to reduce methane production in ruminants. Item 22 provides the use of methane inhibitors and carriers in boluses to reduce methane emissions from ruminants. Item 23 specifies the use of haloform in the manufacture of boluses to reduce the emission of one or more greenhouse gases ("GHGs") from ruminants. Item 24 provides a method for manufacturing any one of items 1 through 18, and this method is a. To form a housing having a cavity, b. Forming a core containing the inhibitor c. This includes transferring the core into the cavity. Item 25 relates to the method of item 24, and the step of forming the core includes mixing the carrier material and the inhibitor. Item 26 provides the method of Item 25, wherein the step of forming the core includes heating the carrier material to melt it before mixing the carrier material with the inhibitor to produce a mixture. Item 27 relates to a method according to any one of items 24 to 26, wherein the step of transferring the core to the cavity includes injecting the mixture into the cavity.

[0142] Further aspects of the present invention should be considered in all its novel forms and will become apparent to those skilled in the art by reading the following description, which provides at least one example of a practical application of the present invention. [Brief explanation of the drawing]

[0143] One or more embodiments of the present invention will be described by reference to the following drawings, not intended to be limiting, but merely illustrative.

[0144] [Figure 1A-B] Figure 1A is a front view of a bolus according to one embodiment of the present invention.

[0145] Figure 1B is a perspective cross-sectional view of the bolus shown in Figure 1A.

[0146] [Figure 2A-B] Figure 2A is a front view of an alternative embodiment of a bolus according to a further aspect of the present invention.

[0147] Figure 2B is a perspective cross-sectional view of the bolus shown in Figure 2A.

[0148] [Figure 3A-B] Figure 3A is a front view of an alternative embodiment of a bolus according to a further aspect of the present invention.

[0149] Figure 3B is a perspective cross-sectional view of the bolus shown in Figure 3A.

[0150] [Figure 4A-B] Figure 4A is a front view of an alternative embodiment of a bolus according to a further aspect of the present invention.

[0151] Figure 4B is a perspective cross-sectional view of the bolus shown in Figure 4A.

[0152] [Figure 5] Figure 5 is a front view of an alternative embodiment of a bolus according to a further aspect of the present invention.

[0153] [Figure 6A-B] Figure 6A is a front cross-sectional view of an alternative embodiment of a bolus according to a further aspect of the present invention.

[0154] Figure 6B is a perspective cross-sectional view of the bolus shown in Figure 6A.

[0155] [Figure 7] Figure 7 is a flowchart showing typical steps of a method for manufacturing a bolus according to one aspect of the present invention.

[0156] [Figure 8-9] Figure 8 is a graph showing the daily diffusion / release rate of bromoform from the bolus in the culture medium.

[0157] Figure 9 is a graph showing the variability in diffusion results.

[0158] [Figure 10-11] Figure 10 is a graph showing the concentration of bromoform in the diffusion medium over time.

[0159] Figure 11 is a graph showing the mass (%) of bromoform released over time.

[0160] [Figure 12-13A] Figure 12 is a graph showing the release rates of bromoform from different carriers in an open-top falcon tube.

[0161] Figure 13A is a graph showing the release rate of bromoform from paraffin wax used as a carrier.

[0162] [Figure 13B-13C] Figure 13B is a graph showing the release rate of bromoform from carnauba wax used as a carrier.

[0163] Figure 13C is a graph showing the release rate of bromoform from beeswax used as a carrier.

[0164] [Figure 14] Figure 14 is a graph showing the average release rate of bromoform for a reinforced bolus according to one embodiment of the present invention.

[0165] [Figure 15A-D] Figure 15A is a side view showing a reinforced bolus design according to an alternative embodiment of the present invention.

[0166] Figure 15B is a side cross-sectional view of a reinforced bolus design according to an alternative embodiment of the present invention.

[0167] Figure 15C is a side cross-sectional view of a reinforced bolus design according to an alternative embodiment of the present invention.

[0168] Figure 15D is a cross-sectional view of the internal structure of a reinforced bolus design according to an alternative embodiment of the present invention.

[0169] [Figure 16A] Figure 16A shows the shrinkage of a tensile test specimen due to injection molding.

[0170] [Figure 16B] Figure 16B shows the absorbed bromoform versus bromoform composition in beeswax for different compositions of PLA mixed with PBS and PBAT.

[0171] [Figure 16C] Figure 16C shows the relationship between absorbed bromoform and the bromoform composition in beeswax for 3D printed PLA and injection-molded 2003D PLA.

[0172] [Figure 16D]Figure 16D shows the bromoform absorption rate versus the bromoform composition in beeswax for different compositions of PLA mixed with PBS and PBAT.

[0173] [Figure 16E] Figure 16E shows the bromoform absorption rate versus PLA composition in beeswax containing different concentrations of bromoform.

[0174] [Figure 17] Figure 17 shows the hardness analysis of the PLA blend before and after exposure to bromoform.

[0175] [Figure 18A-B] Figure 18A shows the release of bromoform from 1 mm thick boluses filled with 67% (by weight) and 55% (by weight) bromoform.

[0176] Figure 18B shows the cumulative release of bromoform from the bolus.

[0177] [Figure 18C-D] Figure 18C shows cumulative plots of 57–1 mm boluses over 7, 8, and 9 days.

[0178] Figure 18D shows the emission velocities from different boluses.

[0179] The term "bromet" used in the diagram refers to bromoform, including bolus. [Modes for carrying out the invention]

[0180] The present invention relates to an apparatus and method for delivering substances, particularly hydrophobic substances, to animals. In a preferred embodiment, the substance is an inhibitor, such as a methane inhibitor. The present invention is illustrated by reference to preferred embodiments. However, this should not be considered to limit the scope of the invention. Those skilled in the art will understand how to apply the teachings herein to apparatuses for delivering other substances to animals.

[0181] First, referring to Figures 1A and 1B, a bolus (100) is provided. The bolus (100) is configured to reduce or eliminate the emission of one or more greenhouse gases ("GHGs") from ruminants. For example, the bolus (100) can reduce or eliminate the production of GHGs by ruminants, and thus reduce the gases emitted by the animals.

[0182] Furthermore, or alternatively, bolus (100) can improve animal productivity by preventing the conversion of feed from ruminants into more than 1 GHG.

[0183] The bolus (100) includes a core (110) and a housing (120).

[0184] In some embodiments, the bolus (100) also includes a barrier layer (130). The barrier layer (130) is configured to separate the core (110) from the housing (120).

[0185] The housing (120) is generally cylindrical and has an open end, generally indicated as (60), and a rounded, closed end (170). The open end (160) allows the fluid in the rumen of the ruminant to come into contact with the core (110).

[0186] Further aspects of Bolas (100) should become clearer from the following discussion.

[0187] core The core (110) comprises at least one inhibitor, which can be optionally mixed with a suitable carrier. Particularly preferred carriers include PEG4000, PEG400, natural and synthetic waxes, fatty acids, aliphatic alcohols, aliphatic amines, phospholipids-lecithin, and adsorbents, as well as combinations thereof.

[0188] Suitable waxes include beeswax, paraffin, castor wax, carnauba wax, candelilla wax, jojoba wax, and lanolin.

[0189] Furthermore, minerals such as zeolite, bentonite, kaolin, activated carbon, or combinations thereof can also be appropriately mixed with the inhibitor. Other compounds such as zinc (i.e., in powder form) or zinc oxide can also be included.

[0190] Alternatively, the core (110) may contain a concentrated (substantially pure) form of the inhibitor.

[0191] In preferred embodiments, the inhibitor is a methane inhibitor. Particularly preferred embodiments include haloforms, such as halomethanes like bromoform (CHBr3)—which will be described in more detail below.

[0192] Those skilled in the art will understand that other carriers can be selected or used depending on the application. It is assumed that specific carriers can be selected to provide a desired release profile for the inhibitor, or to provide a desired physical property of the core material—such as density or volume.

[0193] In a preferred embodiment, the carrier used in the present invention is a natural waxy substance having a preferred melting point of 50 to 90°C, more preferably 60 to 80°C.

[0194] The inventors have found that by having a support having this melting point range, the support can be melted and mixed with the inhibitor, forming a homogeneous core (110), which can then be solidified at room temperature.

[0195] A particularly preferred carrier is a mixture containing castor wax and one or more of paraffin wax, beeswax, and carnauba wax. More preferably, the carrier is a mixture containing castor wax and paraffin wax.

[0196] It should be understood that the carrier-to-inhibitor ratio can be selected to optimize the function of the bolus (100) to match the desired release profile of the inhibitor, for example.

[0197] When formed, the core (containing both the carrier and the inhibitor) preferably has a melting point of at least 45°C. Having this minimum melting point helps ensure that the core (110) does not melt when the bolus (100) is administered to ruminants. Furthermore, it reduces the likelihood that the bolus (100) will melt even if it is inadvertently exposed to high temperatures, such as those that may be reasonably experienced during transport and / or storage.

[0198] It should be understood that the melting point range of the core (110) can be adapted by changing the ratio of the inhibitor to the carrier forming the core (110).

[0199] Preferred ratios of inhibitor to carrier include substantially 80:20 w / w% to substantially 50:50 w / w%, preferably substantially 70:30 w / w% to substantially 60:40 w / w%, or more preferably substantially 66:33 w / w%.

[0200] inhibitors In a preferred embodiment, the inhibitor is one or more methane inhibitory compounds.

[0201] Suitable methane inhibitors include haloforms such as bromoform, chloroform, and iodoform, and combinations thereof. Any methane inhibitor suitable for internal administration to ruminants is expected to be usable in this invention.

[0202] The inventors have surprisingly found that bromoform is particularly well suited for use in the bolus (100) according to the present invention. Therefore, the inhibitor is referred to as bromoform in this specification. However, this should not be considered to limit the scope of the invention, and alternative examples are also considered to be within the scope of the invention.

[0203] Bromoform is reactive and has a short half-life in animals (0.8 hours in rats, 1.2 hours in mice, U.S. Department of Health and Human Services, 2003). It is a liquid at room temperature and is denser than water. Previous studies have demonstrated that no residue is present in the meat or tissue of slaughtered castrated cattle even after 48 hours, including storage time (Kinley et al. Mitigating the carbon footprint and Improvement Productivity of ruminant livestock Agriculture using a red foam, Journal of Cleaner Production 259 (2020) 120836), and there is no significant increase in levels in milk (Roque et al. Inclusion of Asparagopsis armata in lactionating pressures' Diet is enteric methane exit by over 50%; Journal of Cleaner Production 234 (2019) 132-138).

[0204] Bromoform has relatively high efficacy, for example, an effect per dose. This makes it possible to provide within the core (110) a sufficient amount to produce a bolus (100) that can deliver a controlled release of the inhibitor over a long period of time.

[0205] Additionally, bromoform is relatively dense. This can help achieve higher retention of the bolus (100) within the lumen by optimizing the density of the bolus to encourage it to sink into the ventral portion of the lumen rather than float.

[0206] Despite the points mentioned above, there are general concerns about the use of bromoform in animals. This compound is thought to have adverse effects, including carcinogenicity, at certain exposure levels.

[0207] Furthermore, there are technical challenges when administering bromoform to animals. These include the volatility of the substance and its ability to dissolve the substances used for its delivery. In addition, achieving a precise (and relatively low) administration rate over a period of time is a challenge.

[0208] housing The housing (120) includes a cavity (not numbered in the diagram) of a size and dimensions that accommodates the core (110). The housing (120) forms the external structure of the bolus (100).

[0209] The housing (120) is configured to provide structural integrity to the bolus (100), but is also adapted to degrade over time. Degradation of the housing (120) may facilitate the release of the inhibitor over a given period of time.

[0210] The housing (120) is preferably non-toxic and withstands erosion within the rumen of the ruminant for a period sufficient to facilitate the release of the inhibitor from the core (110) at a desired rate. It should be understood by those skilled in the art that the dissolution rates of the housing (120) and the core (110) may be configured to allow for controlled release of the inhibitor within the rumen of the ruminant.

[0211] Preferably, the housing (120) is made of a biodegradable, non-absorbable material, or a material suitable for waste disposal in a slaughterhouse. It should be understood that any material suitable for internal administration to ruminants at a desired dissolution rate can be used in this invention.

[0212] In preferred embodiments, the housing (120) is preferably selected from biodegradable materials, particularly preferably from biodegradable materials including polymers such as polylactic acid (PLA), polyglycolic acid (PGA), polyglycolic acid polylactic acid (PLGA), polypropylene, SLA polymer, PBS, and combinations thereof. In particularly preferred embodiments, the housing (120) is made of a material containing PLA and PBAT.

[0213] In a preferred embodiment, the housing (120) is made of PLA. PLA is available in three forms: D-, L-, and racemic mixtures of both D and L. All three types of PLA can be used in the housing (120) of the present invention.

[0214] In its preferred form, PLA that decomposes into lactic acid is preferred and is commonly used as a medical implant. Depending on the type of PLA used, it decomposes in the body within 6 months to 2 years.

[0215] Those skilled in the art should understand that other suitable biodegradable materials can be used as the housing (120).

[0216] In any embodiment, further fillers, binders, surfactants, activators and / or absorbents may be included in the bolus of the present invention.

[0217] As shown in Figures 1A and 1B, the bolus (100) has a substantially cylindrical shape. The housing (120) includes a smooth outer surface to facilitate ingestion of the bolus (100) by ruminants.

[0218] Those skilled in the art will understand that the size, thickness, and / or dimensions of the bolus (100), including the core (110), the barrier layer (130) if provided, and the housing (120), can be adjusted according to the dosage of the inhibitor delivered to the ruminant, without departing from the spirit and scope of the invention. For example, smaller sized bolus (100) can be adapted for use in smaller ruminants such as sheep and goats, and larger sized bolus (100) can be used in larger ruminants such as cattle. A bolus for a large animal such as a cattle may have dimensions of 13 cm in length, 3.4 cm in diameter, and weigh 257 gm (throughout this specification, "gm" refers to grams). A bolus for a relatively small animal such as a sheep may have dimensions of 8.5 cm in length, 2 cm in diameter, and weigh 60 g. Alternatively, smaller boluses can be administered to relatively large ruminants such as cattle, and such smaller boluses may have dimensions of 3.4–3.8 cm in length and 2.6–3.0 cm in diameter.

[0219] This paper also considers the possibility of using multiple smaller boluses in combination. In preferred embodiments, the bolus and retarder formulation of the present invention have a length of at least 5 cm, most preferably at least 10 cm, and more preferably 10.3 cm. In preferred embodiments, the bolus and retarder formulation of the present invention have a diameter of at least 2 cm, preferably 3.4 cm, and a length of at least 10 cm, preferably 10.3 cm. Preferably, the bolus and retarder formulation of the present invention have a weight of 100 to 300 grams.

[0220] Additionally, the housing (120) may be configured to control the release rate of the core (110) and / or the decomposition of the bolus (100). For example, the internal cross-sectional area of ​​the cavity may be adapted to control the amount of core (110) present within the bolus (100). In such embodiments, the internal volume of the cavity may be adapted to increase in size from the open end (160) to the closed end (170). This may help to increase the amount of inhibitor over time. This may explain the growth of animals as their feed intake increases.

[0221] Additionally, or alternatively, the cross-sectional thickness of the walls forming the housing (120) can be increased along the length of the housing (120). For example, the walls may be thicker at one end of the housing (120) than at the other end. In such embodiments, the thickness of the walls at the open end (160) may be thinner than that towards the closed end (170). This helps to provide controlled dissolution of the core formulation from the bolus.

[0222] Barrier layer The barrier layer (130) is an optional component of the bolus (100) of the present invention and may be included to provide further stability to the bolus (100). The barrier layer (130) may be configured to partially or completely prevent contact between the core (110) and the housing (120). The barrier layer (130) is preferably selected from a waxy material, epoxy, or silicone material.

[0223] Those skilled in the art will understand that the barrier (130) layer can be selected according to the desired application and / or release profile. For example, if further control of the inhibitor release rate is desired, the desired release profile can be easily obtained by selecting the material, shape, and configuration of the barrier layer (130).

[0224] Composition example In an exemplary embodiment, the bolus may comprise a core enclosed by a housing. The bolus may be approximately 13 cm in length, 3.4 cm in diameter, and weigh approximately 257 g.

[0225] The housing is made of PLA (such as 3052D, 3001D, 3251D, L130, etc.) and may be manufactured by injection molding, for example, with a thickness of 1 mm.

[0226] The core matrix can be prepared from a mixture of castor wax and paraffin wax in a 50:50 (by weight) ratio. This matrix may contain bromoform as an inhibitor at a concentration of approximately 50% (by weight).

[0227] Treatment method The bolus (100) is delivered orally into the rumen of the ruminant being treated and enters the rumen via the esophagus. Within the rumen, gastric juices (and other substances such as plant fiber mat) act to eventually erode or dissolve the core (110), releasing the inhibitor over time. However, the housing remains substantially intact during the treatment period.

[0228] The open end (160) allows gastric juice and fibrous material to come into contact with the core (110). Furthermore, it helps to control the release of the core (110) from there into the lumen.

[0229] The core (110) and housing (120) are designed to facilitate the release of the inhibitor over the period during which the animal is treated according to the methods disclosed herein.

[0230] The bolus (100) is adapted to release the inhibitor over a period of at least 6 months, preferably 12 months, and possibly up to 2 years.

[0231] Preferably, the inhibitor release rate can be calculated based on the body weight of the ruminant being treated and the type of inhibitor used. Therefore, it will be understood that the desired release rate may vary from animal to animal. Typically, the desired release rate can be calculated based on the amount of inhibitor / animal body weight. Alternatively, the desired release rate can also be calculated based on the amount of feed consumed by the animal. Particularly preferred release rates for bromoform include about 0.1 to about 0.5 g / day, more preferably about 0.2 g / day.

[0232] Additionally, those skilled in the art should understand that ruminants can be treated by multiple boluses (100) according to the present invention to achieve a preferred dose of the inhibitor. This makes it possible to produce boluses (100) having concentrations and total amounts of the inhibitor. Multiple of these boluses (100) can be administered to the animal simultaneously or sequentially. This makes it possible to deliver the desired dose to the animal. This is particularly useful for animals requiring different doses of the inhibitor, for example, larger or smaller animals, or to supplement natural growth over time.

[0233] A bolus (100) is adapted to deliver a certain dose of the inhibitor directly into the animal's rumen. For example, bromoform may be released at a rate that effectively reduces or eliminates methane production during digestion. This reduces greenhouse gas emissions from animals and also reduces the environmental impact of agriculture.

[0234] Furthermore, the bolus (100) may improve the conversion of feed for animal production in ruminants. For example, by reducing methane production during digestion, it is thought that this could lead to more efficient utilization of ingested feed, potentially resulting in improvements in growth, weight gain, or other production such as milk production. In addition, the synergistic effects resulting from the core composition and the combination of carrier and inhibitor may enable the provision of sustained-release, long-term delivery devices to improve animal productivity and / or reduce greenhouse gas emissions.

[0235] First Alternative Housing Embodiment Referring now to Figures 2A and 2B, an alternative embodiment of the bolus (200) according to one embodiment of the present invention is shown.

[0236] The aspects of bolus (200) are the same as those of bolus (100), and therefore similar reference numerals refer to similar components.

[0237] A series of ribs (240) are provided along the outer surface of the housing (120). The ribs (240) can provide additional structural strength to the bolus (200) and can help prevent the bolus (200) from rupturing if the core (110) expands. Additionally, or alternatively, the ribs (240) can also assist in the administration of the bolus (200) to ruminants.

[0238] As shown in the illustration, the ribs (240) are provided as a series of concentric "hoops". However, the ribs (240) may be a series of parallel or non-parallel ribs (not shown) extending along the length of the bolus (200).

[0239] Second Alternative Housing Embodiment Next, we refer to Figures 3A to 3B, which show an alternative embodiment of the bolus (300) according to one embodiment of the present invention.

[0240] The aspect of bolus (300) is the same as the aspect of bolus (100) described above, and therefore similar reference numerals refer to similar components.

[0241] The bolus (300) includes additional features on the outer surface of the housing (120), including a recess or groove (350).

[0242] The groove (350) can facilitate the detachment of a portion of the housing (120) as the housing (120) decomposes. This can be used to further control the release profile of the inhibitor.

[0243] Third Alternative Housing Embodiment Next, refer to Figures 4A to 4B, which show an alternative embodiment of the bolus (400) according to one embodiment of the present invention.

[0244] The aspect of bolus (400) is the same as the aspect of bolus (100) described above, and therefore similar reference numerals refer to similar components.

[0245] The bolus (400) includes a housing (120) having a cavity (not shown) configured to receive and hold a core (110).

[0246] The housing (120) is tapered along its length. For example, the distance between the distal outer surfaces of the housing (120) increases along the length of the bolus (400). For example, as shown in Figure 4A, the width (X) is smaller than the width (Y).

[0247] Alternatively, the bolus (400) may have sidewalls of substantially constant thickness, but is structured and oriented to define the taper of the bolus (400).

[0248] This configuration allows for better control over the degradation of the core (110), thereby enabling further control over the release of the inhibitor.

[0249] Fourth Alternative Housing Embodiment Referring now to Figure 5A, an alternative embodiment of the bolus (500) according to one embodiment of the present invention is shown.

[0250] The bolus (500) is similar to that described above, and therefore similar reference numerals refer to similar components.

[0251] The bolus (500) comprises a reservoir (580) adapted to hold a relatively concentrated form of the inhibitor, such as bromoform in a substantially pure liquid form.

[0252] The bolus (500) includes a dispensing mechanism configured to dispense a predetermined dose of the inhibitor from the reservoir (580).

[0253] In the illustrated embodiment, the dispensing mechanism is a pump (590) in communication with a valve. At a predetermined time, the pump (590) dispenses a dose of the inhibitor through the valve (590), and a bolus (500) releases the inhibitor into the lumen to which it was administered.

[0254] The dispensing mechanism may be configured to release the same amount of inhibitor at a specified interval, for example, due to its consistency.

[0255] Alternatively, the distribution mechanism may be configured to vary the amount of inhibitor released at different time points. This may be useful in ensuring that a bolus (500) provides an effective dose of the inhibitor that takes into account animal growth. Furthermore, or instead, it may also compensate for changes in other factors, such as seasonal variations in methane production requiring larger doses of inhibitor.

[0256] In further embodiments, the bolus (500) may include sensors (not shown). For example, a temperature sensor may be included within the bolus (500). Additionally or alternatively, other sensors such as motility or pH may also be included in the bolus. Adding such sensors provides valuable information about the animals' feed intake and allows for assessment of whether the amount of inhibitor is sufficient for the animals.

[0257] Fifth Alternative Housing Embodiment Next, we refer to Figures 6A and 6B, which show an alternative embodiment of the bolus (600) according to one embodiment of the present invention.

[0258] The bolus (600) may be adapted to include additional features within the cavity of the housing, such as grooves or ribs (680) formed in the inner wall of the housing (120) that define the cavity.

[0259] The embodiment of the bolus (600) is similar to the embodiment of the bolus (100), and thus, like reference numerals refer to like components.

[0260] A series of ribs (680) are provided along the inner surface of the housing (120). The ribs (680) can provide additional structural strength to the bolus (600) and / or provide additional means for retaining the contents of the core formulation within the cavity of the housing. Additionally or alternatively, the ribs (680) can also assist in retaining the core within the housing. Further, the ribs can also provide for the controlled dissolution of the core formation from the bolus (600) into the ruminant.

[0261] In one embodiment, the outer surface of the housing remains smooth or uniform.

[0262] Sixth Alternative Housing Embodiment Next, refer to FIGS. 15A - 15D showing further embodiments of a bolus (700) according to one aspect of the present invention. The dimensions of the bolus in the figures are shown in mm. Preferably, the bolus has a length of 13 cm, a diameter of 3.4 cm, and preferably weighs approximately 250 grams.

[0263] The bolus (700) can be adapted to include additional features with an internal reinforcement structure on the housing.

[0264] The embodiment of the bolus (700) is similar to the embodiment of the bolus (100), and thus, like reference numerals refer to like components.

[0265] The bolus (700) includes at least one reinforcing rib (710) located within a cavity (unnumbered) defined by the housing structure. For example, a cap (720) may also be provided, which is releasably attached to the bolus (700) to close the open end of the bolus (700). The attachment may be provided by a friction fit configuration or by a threaded configuration in which corresponding threads on the housing and the cap engage with each other. Alternatively, the cap may be attached to the housing by adhesive or other mechanical fasteners.

[0266] The reinforcing rib (720) can improve the structural integrity of the bolus (700) and help the bolus maintain its shape.

[0267] Manufacturing method Referring now to Figure 7, this is a flowchart showing typical steps in the manufacturing method (800) of boluses (100), (200), (300), and (400) according to the present invention, for example.

[0268] Generally speaking, this method includes the steps of forming a housing (120) (810) and forming a core (110) (820).

[0269] housing The housing (120) can be formed using any technique known to those skilled in the art. For example, a suitable material can be extruded into the desired shape to define the cavity. Alternatively, an additional additive manufacturing process can be used to construct the housing shape defining the cavity. It is also conceivable that a molding process may be used, which may include sacrificial molding or injection molding, 3D printing or hot melt extrusion.

[0270] core In step 820, the core (110) is manufactured.

[0271] Step 820 can include one or more of the following steps.

[0272] Step 822 includes melting the carrier material to provide a melted carrier material.

[0273] Step 824 includes adding an inhibitor to the melted carrier material.

[0274] In step 826, the inhibitor and the melted carrier material are mixed to produce a substantially homogeneous mixture.

[0275] Step 828 includes forming the substantially homogeneous mixture into a desired shape.

[0276] It should be understood that the substantially homogeneous mixture contains the inhibitor at a concentration sufficient to achieve the desired release profile of the inhibitor upon administration of the device to a ruminant. The concentration can be varied depending on the type of ruminant to be treated, the shape and dimensions of the device, or the desired release profile to be achieved.

[0277] It should be understood that the step of forming the substantially homogeneous mixture into a desired shape may include feeding the mixture into a mold. In a particularly preferred form, the substantially homogeneous mixture is added (injected) into the cavity within the housing (120) manufactured in step 810.

[0278] Alternatively, the mold may be a separate component that receives the substantially homogeneous mixture. In these embodiments, once the desired shape is formed, a core can subsequently be provided in the cavity within the housing (120).

[0279] This method also includes the step of allowing the substantially homogeneous mixture to cool. As it cools, the carrier material hardens and takes the shape corresponding to the shape of the mold or housing in which it is incorporated.

[0280] Example formulation The following cores were formulated for use in the bolus of the present invention. [Table 2] [Table 3] [Table 4] [Table 5]

[0281] verification Example 1: Release / Diffusion Study A test (RME Test 2) was conducted using a 2mm thick, 3D-printed large capped bolus (LCB2) filled with 66.7% (by weight) bromoform and 33.3% (by weight) beeswax inside a Lumen Emulator (RME) to determine the diffusion rate of bromoform from the bolus.

[0282] Bolas Design In this study, a reinforced bolus, as shown in Figure 15, was used. It included an internal reinforcing structure and widened ribs to support the walls, and the top was fitted for attaching a cap. The reinforced bolus was found to be more robust and better retain its shape than the unreinforced bolus when injected with molten bromoform / beeswax mixture and cooled, resulting in a physically more robust bolus in the tests.

[0283] method material Bromoform (reagent grade, Sigma Aldrich, 96% bromoform, 4% ethanol), beeswax (food grade, NZ beeswax, MP65℃), and zinc oxide from Native Elements NZ.

[0284] Bolus manufacturing The bolus was drawn in Solidworks, converted to an .stl file, opened in FlashPrint, and a print job was created. The bolus was printed in three parts (case, internal structure, and cap) on a FlashForge Creator Pro 3D printer using 100% filled E-Sun PLA+, at standard resolution, first layer height 0.27mm, layer height 0.18mm, 2 outer shell layers, 3 upper solid layers, 3 lower solid layers, hexagonal filling pattern, print speed 60mm / s, extruder temperature 200°C, and plate temperature 50°C.

[0285] Eight LRB boluses were prepared with 67% (by weight) bromoform, eight LRB boluses were prepared with 75% (by weight) bromoform, and six LCB2 boluses were prepared without bromoform (control). The components are listed below (Table 1). All components were weighed in beakers using a calibrated 4dp electronic balance. The bromoform solutions were covered with Parafilm to prevent evaporation. The components were prepared by melting pre-weighed beeswax and zinc oxide in a beaker at 100°C (thermoprism oven), cooling the mixture to 80°C, adding bromoform, and thoroughly mixing the mixture to prevent zinc oxide sedimentation before injecting it into the boluses. The boluses were sealed by press-fitting and soldering caps. [Table 6]

[0286] The bolus was placed in a 500 ml polypropylene bottle with approximately 380 ml of 0.02 M phosphate buffer (Merck) in distilled water, and prepared in batches of 2 L or more. The pH was adjusted to 6.5 using 1 M HCl (Merck) and a pre-calibrated pH meter (using pH 4, 7, and 10 pH buffers). The bottles were sealed and placed in a 40°C incubator. 10 ml samples were collected, and the entire solution was replaced every 24 hours.

[0287] 10 ml of sample was collected into 15 ml Falcon tubes using a 10 ml autopipette. 1 g of sodium chloride was added to each Falcon tube. For GC-MS analysis, 1 ml of ethyl acetate (analytical grade, Merck) was added to each Falcon tube. If using GC-FID, 2 ml of ethyl acetate was added to each Falcon tube. The Falcon tubes were capped, thoroughly mixed using a vortex, and centrifuged at 4000 rpm for 15 minutes. For GC-MS analysis, all ethyl acetate was collected using a graduated glass syringe and the volume was recorded.

[0288] For GC-FID analysis, 0.5 ml of ethyl acetate was recovered. For GC-FID analysis, 200 μl of the sample was injected using an autosampler, and the analysis was performed in splitless mode using a ZB5HT30m capillary column with a temperature gradient of 30–300°C over 20 minutes under a nitrogen gas flow of 5 ml / min. The retention time for bromoform was 7.5 minutes. The mass (mg) of bromoform was determined by comparing the peak area to a calibration standard prepared with ethylene acetate. This was divided by the injection volume to determine the concentration of bromoform in ethyl acetate (mg / L). The concentration in ethyl acetate was multiplied by the total amount of ethyl acetate added to the sample and divided by the recovery rate to determine the mass of bromoform in the sample. This was then divided by the volume of the collected sample to determine the concentration in the solution, and this was then multiplied by the volume of the solution in the shot bottle to determine the mass transferred from the bolus to the solution. The bromoform recovery rate from the solution was checked using standard solutions prepared with bromoform at various concentrations, and was typically 43%. The performance of the GC-FID was checked every 10 sample runs, using a calibration sample as a reference.

[0289] result Both bolus applications resulted in a low diffusion rate followed by a rapid increase in diffusion rate (Figure 8). The 67% bolus required a delay of 4–5 days to reach the maximum diffusion rate, while the 75% bolus reached the maximum diffusion rate in 3 days.

[0290] The diffusion rate was higher with the 1010 mg / day bolus (75%) compared to 66.7% with the 730 mg / day bolus. This was surprising, but also a good result, as the predicted diffusion rate for the 67% bromoform LCB1 bolus was 300 mg / day, and for the 75% bromoform LCB1 bolus it was 462 mg / day (meaning that methane reduction can be achieved by administering a single bolus to a 700 kg bull). A lower diffusion rate was expected for the LRB bolus due to its reduced surface area (1 mm thick, approximately 71% of the LCB1 bolus) (Table 2). Theoretically, the LRB bolus should only need to deliver 220 mg / day for 67% bromoform and 344 mg / day for 75% bromoform. [Table 7] [Table 8]

[0291] The variability in the diffusion data initially had a high coefficient of variation of approximately 1, which decreased to 0.05-0.22 as the bolus reached its maximum diffusion rate (Figure 9). The 75% bolus stabilized within 2 days, and the 67% bolus stabilized within 4 days.

[0292] Zero-order emission was observed in both boluses, indicating that the emission rate was independent of the bromoform concentration in the bolus (Figure 11).

[0293] conclusion The diffusion rates of LRB boluses were 1010 mg / day for 75% boluses and 730 mg / day for 66.7% boluses, which were higher than predicted from previous diffusion studies.

[0294] Since the concentration of bromoform in a 75% bolus of culture medium is close to the solubility limit of bromoform in water (3.2 g / L), the diffusion rate may be higher than that measured in this study.

[0295] Example 2: Carrier release test For this study, release tests of various carriers were conducted.

[0296] method material Bromoform (reagent grade, Sigma Aldrich, 96% bromoform, 4% ethanol), rumen solution (Dairy NZ test), paraffin wax (MPs46-48, 55 and 65°C, Sigma Aldrich), castor wax (Lotus Oils), carnauba wax (PureNature NZ), zinc oxide (PureNature NZ).

[0297] pH and buffering capacity of rumen fluid Rumen fluid collected from Dairy NZ was thawed, centrifuged, and then analyzed for pH and buffering capacity. 10 ml of rumen fluid was taken from each cow and titrated with 0.05 N NaOH while continuously monitoring the pH. The amount of NaOH required to change the pH in units was recorded.

[0298] Release and testing of various carriers As described in Example 1 above, a small bolus with a cap was prepared.

[0299] Paraffin wax, beeswax, carnauba wax, and castor wax were mixed with bromoform to achieve bromoform content of 33%, 50%, 67%, and 75% by weight. The mixes were arranged as follows: a. Paraffin wax: 2mm thick small capped bolus and 15ml falcon tube; b. Castor oil, carnauba wax, and beeswax: 1, 2, and 3 mm boluses with small caps and 15 ml falcon tubes.

[0300] These were placed in 500 ml polypropylene bottles with 400 ml of 0.02 M phosphate buffer (Merck) in distilled water, and prepared in batches of 2 L or more. The pH was adjusted to 6.5 using 1 M HCl (Merck) and a pre-calibrated pH meter (using pH 4, 7, and 10 pH buffers). The bottles were sealed and placed in a 40°C incubator. 10 ml samples were collected, and the entire solution was replaced every two days (Monday, Wednesday, and Friday), excluding weekends.

[0301] As described in Example 1 above, the samples were analyzed by GC-MS and GC-FID.

[0302] result pH and buffering capacity The average pH and buffering capacity were 6.9±0.2 (n=4) and 7.47±1.4 mMol / L / DeltapH (n=4), respectively. While literature on the pH value of rumen fluid has been published, data on buffering capacity was not available. The buffering capacity obtained for rumen fluid is 5-6 times higher than that of phosphate-buffered saline, indicating that the rumen environment is resilient. The pH of the phosphate buffer in the diffusion experiment was found to be stable even at a bromoform concentration of around 3 mg / ml (Report No. BR2021-01, Figure 4). Assuming a rumen fluid volume of 91 L, the maximum bromoform concentration in the extreme condition of a complete bolus rupture reached approximately 1.09 mg / ml, which was lower than previously observed in PBS. Therefore, considering the strong buffering capacity of the rumen fluid at this concentration, the likelihood of a decrease in pH even in the case of a sudden bolus rupture is low.

[0303] Carrier release test The release rate of paraffin wax was highest at 190 mg / cm² / day, followed by beeswax, carnauba wax, and castor wax (Figure 12). Carnauba wax and castor wax appear to be better carrier options because their release rates are 50-40% lower than those of beeswax.

[0304] Bromoform showed the maximum release rate of 3.5–5.4 mg / cm² / day in small, capped boluses made of paraffin wax (Figures 13A–C).

[0305] The release rates of boluses made from carnauba wax were a maximum of 5.5 mg / cm² / day for a bolus with a thickness of 1 mm and 1.66 mg / cm² / day for a bolus with a thickness of 3 mm.

[0306] In comparison, a bolus made from beeswax showed a release rate of 3 mg / cm² / day with 75% (by weight) bromoform (Figure 13C).

[0307] Bromoform dissolved the castor wax, diffused through the bolus, accumulated at the bottom of the container, dissolved the container, and since no bromoform was detected in the collected water sample, the release rate could not be determined. The castor wax experiment could be repeated in a glass bottle.

[0308] Enhanced discharge rate from Bolas From another test, Figure 14 shows the average release rates of large reinforced boluses containing 67% (by weight) and 75% (by weight) bromoform, prepared as described in Example 1 above, and compared them to the release rates from the same boluses measured in the laboratory. Half of the boluses were placed in a 20L bucket containing 1 kg of sand filled with pH 6.5 buffer, and the other half were placed in a 20L bucket containing 400 g of wood chips and 1 kg of sand. The release rates on day 28 were comparable to those observed in the laboratory. There was little difference in bromoform concentration between the buckets with and without wood chips. The boluses remained mostly intact, but were compressed by the sand, and some had their lids open.

[0309] Example 3: Animal Research Animal studies were conducted to measure methane emissions from animals implanted with the bolus of the present invention. This experiment was designed as an unbalanced and fully randomized design, involving three treatments at three intervals separated by 8–12 weeks and three repeated measurements over time.

[0310] From a research farm in Manawatu, New Zealand, 19 heifers (live weight 312±14kg), including 3 reserves, were selected from a population of 50 based on behavioral characteristics and live weight. These were assigned to one of three treatments: a bolus without bromoform (control; n=4); a bolus releasing bromoform at a rate of approximately 300–400 mg / day (low, n=6); or a bolus releasing approximately 450–580 mg / day (high, n=6). SmaXtec boluses were administered concurrently to monitor rumen temperature as an animal health monitor and to supplement weekly blood samples.

[0311] Heifers were transported from the research farm to the test center for dietary adaptation and gas measurements using a respiratory chamber. The heifers were adapted to the barn and freshly mowed pasture environment for seven days before receiving their assigned therapeutic bolus. Gas measurements began 13 days after bolus administration. During the gas measurement period, each heifer was placed in a respiratory chamber for 48 hours, a process that took two weeks across four measurement groups. Once the respiratory chamber measurements were complete, the animals were returned to the research farm.

[0312] Bolas preparation The bolus was prepared according to the procedure described in Example 1 above. The following formulations used in this test are shown in Table 4 below. [Table 9]

[0313] Bolus administration Three versions of the bolus were administered within the first 10 days of the experiment. The first version was a short bolus that all animals regurgitated within 5 days of administration. The control bolus was longer than the treatment bolus and did not regurgitate for the first 3 days, so it was assumed that bolus size was the main factor in regurgitation. All first-version treatment boluses were replaced with second-version boluses on day 5 after administration. However, the longer boluses of the second version also regurgitated. Therefore, these boluses were replaced with a third-version treatment bolus, which was a significantly heavier bolus of the same size as the second-version boluses. The third-version boluses have not regurgitated to date. Currently, almost all heifers have been administered the third-version bolus, with the exception of three low-treatment heifers. Details of bolus regurgitation and re-administration are shown in Table 5.

[0314] Two control boluses were refluxed, but only one was identified because the bolus ID could not be deciphered. Neither control bolus was re-administered because it was not possible to determine if the heifer was a match for the bolus. [Table 10]

[0315] Feed intake and live weight Heifers that had not yet calved were given free access to cut ryegrass-based pasture. The feed was harvested at the research farm at approximately 10:00 AM daily and transported to the test center. The harvested feed was divided into two portions; the first portion was fed at 3:30 PM, and the second portion was stored at 4°C until 8:30 AM the following morning. Samples were collected from each grass supply for dry matter measurement and feed analysis. Dry matter (DM) was determined by oven drying three subsamples at 105°C for 24 hours. For chemotonutrient analysis, another subsample was oven dried at 65°C for 48 hours. Both drying ovens used were forced-air ovens (Avantgarde FED 720, Binder GmbH, Germany).

[0316] Two days prior to entering the respiratory chamber for methane measurement, the cows were placed in a metabolic box and restrained to allow them to adapt to the confined space. While the animals were in the metabolic crate or respiratory chamber, rejected feed was collected twice daily, and the rejected DM was measured as described above. The daily dry matter intake of heifers was then determined from the difference between the offered dry matter and the rejected dry matter.

[0317] Live weight was recorded twice (July 13, 2021 and July 16, 2021) while the animals were grazing at the research farm prior to the experiment. Animal weight was measured again upon arrival at the test farm on July 19, 2021, and every 7–10 days while in the field. The initial live weight was measured on July 23, 2021, before bolus administration, and the final live weight was measured after the animals were removed from the breathing chamber. Because measurements were taken over a two-week period, the final live weight dates differ for some animals.

[0318] Gas measurement The fermentation gases, methane (CH4), carbon dioxide (CO2), and hydrogen (H2), were quantified in four open-circuit breathing chambers at the New Zealand Ruminant Methane Measurement Centre (AgResearch, Palmerston North, New Zealand). Each chamber measured 15.4 m³. 3 (Length 3.5m x Width 2m x Height 2.2m), airflow velocity is approximately 1.0m 3The differential pressure was continuously monitored at a rate of / min by measuring it using a Venturi flow meter. The temperature inside the breathing chamber was approximately 20°C, and the relative humidity averaged approximately 79%. All gases were measured at approximately 2.8-minute intervals using a 4900C Continuous Emission Analyzer (Servomex Group Ltd, East Sussex, UK), and the daily production of each gas was calculated from the difference between the concentration entering and leaving the chamber (Pinares-Patino et al., 2012). The breathing chamber was opened twice a day (for approximately 20 minutes each time) for cleaning, feeding, fecal sampling, and collection of feed refusal. No measurements were taken while the chamber was open, and missing data was interpolated by taking the average of the last 12 values ​​(approximately 45 minutes) before the door was opened.

[0319] statistical analysis Gas measurement data from the initial period were analyzed using the "predictmeans" and "lme4" packages in the statistical software R4.0.3 (R Core Team, 2020). Dry matter intake and gas emissions data for each heifer were averaged over two measurement days. Heifers served as the experimental unit. The mixed model included treatment as a fixed effect and a respiratory chamber nested in the measurement group as a random effect.

[0320] Live weight analysis included treatment as a fixed effect and time as a repeated measure, with heifers being used as the subject of the repeated measure. This analysis included only the initial and final live weights.

[0321] result Dry food intake and gas emissions Administration of bromoform at approximately 300-400 mg / day (low) or approximately 450-580 mg / day (high) to heifers did not affect dry matter intake measured during the two days spent in the respiratory chamber compared to the control group (p=0.42). Both: CH4 production (g / day) and CH4 yield (g / kg dry matter intake units) were reduced by more than 99% in both the low and high treatments compared to the control (p<0.01). The reduction in CH4 emissions in both low and high treatments was accompanied by an increase in H2 emissions per day (Table 7). Since methane emissions were completely reduced in both treatments, methane reduction levels of 30-90% can be achieved with lower doses. Reducing the daily dose prevents the use of more bromoform than necessary to extend the lifespan of the bolus, reducing the risk of adverse effects on animals and potential contamination of animal products. Given that methane excretion was completely suppressed, it is noteworthy that there was no adverse effect on dry matter intake, as observed when bromoform containing asparagopsis was administered (Roque et al. 2019). [Table 11]

[0322] conclusion As observed, the above results demonstrate that the treatment using the bolus according to the present invention is highly effective within several weeks after bolus administration, as evidenced by the approximately 99% reduction in methane.

[0323] Unless the context clearly requires otherwise, terms such as “includes” and “contains” throughout the description and claims should be interpreted in a comprehensive rather than exclusive or exhaustive sense, meaning “includes but not limited to.”

[0324] All applications, patents and publications cited above and below are incorporated herein by reference, if any.

[0325] References to prior art in this specification do not constitute, and should not be interpreted as, an endorsement or any suggestion that such prior art forms part of common general knowledge in any field of effort in any country in the world.

[0326] The present invention can also be broadly said to consist of parts, elements, and features that are individually or collectively referred to or shown in the specification of this application, and any or all combinations of two or more such parts, elements, or features.

[0327] Wherever a component having an integer or a known equivalent is referred to in the preceding description, those integers are incorporated herein as if they were described separately.

[0328] It should be noted that various changes and modifications to the currently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the invention and without impairing the associated advantages. Therefore, such changes and modifications are intended to be included in the invention.

[0329] Example 4 method material PLA(3052D), PBS (supplier Convex), and PBAT (supplier Convex) were freeze-dried in aluminum foil trays using a Labcono freeze-dryer before use to reduce the moisture content in the blend.

[0330] Bolus manufacturing The blend of PLA(3052D), PBS, and PBAT was prepared by mixing the pellets in the following ratios. [Table 12]

[0331] The blend was prepared by melt blending in a LabTech co-rotating twin-screw extruder (L / D 44:1) at a screw speed of 200 rpm. The temperature profile rose across 11 barrel heating sections, from 70°C at the feed throat to 220°C along the main barrel and up to 230°C at the die. The blend was granulated using a tri-blade granulator with 4 mm plates (Castin Machinery, NZ). The blend was stored in aluminum foil trays and packed into Ziploc® bags before use. All blends were oven-dried overnight at 40°C before injection molding. Tensile (ASTM D368) and impact (ISO 179) specimens were produced in a BOY 35A injection molding machine with a temperature profile of 70–220°C from feed to nozzle. The mold temperature was kept constant at 50°C. Lanolin was used as a release agent and sprayed onto the mold before each tensile bar was produced.

[0332] Bolas's analysis Shrinkage due to injection molding was determined by measuring the width and thickness of the tensile test specimen, subtracting this from the width and depth of the mold, dividing by the width and depth of the mold, and multiplying by 100 to obtain a percentage. The tensile rod was cut to a length of approximately 2 cm using a band saw, and the ends were polished smooth using 500 grit sandpaper. A flat-bottomed glass petri dish with a diameter of 120 cm was filled with the following bromoform concentrations: beeswax / bromoform mixtures of 33, 50, 67, and 75 wt%. Three samples of each PLA blend were labeled, weighed on a 4dp electronic balance, and measured in thickness, length, and width using digital calipers. These were then laid flat and slowly pressed into each bromoform / beeswax mixture to ensure good contact between the beeswax and the PLA surface. The glass lid was then placed on top of the petri dish, sealed with insulating tape, and placed in a 40°C incubator.

[0333] The hardness of the sample was tested using a Shore D hardness tester with a weight of 7 kg, and its structural properties were also tested using XRD.

[0334] Every two or three days, the sample was removed from the petri dish, washed with tissue paper, weighed using a 4dp electronic balance, and measured using digital calipers.

[0335] The absorption of bromoform was determined by measuring the total change in sample mass and dividing it by the sample's starting mass. The absorption rate was determined by dividing the change in sample mass between measurements by the area of ​​the sample in contact with the bromoform / beeswax mixture and then dividing that by the change in time between measurements.

[0336] Swelling was determined by measuring the change in the sample's volume and dividing it by the sample's original volume.

[0337] result injection molding The shrinkage of the PLA was approximately 0.2%, but increased to approximately 1–1.2% with increasing amounts of the PBS and PBAT blend (Figure 16). A person skilled in the art would know how to adjust the shrinkage to produce boluses of desired size and dimensions. It should be understood that while various sizes of boluses are possible, given the teachings of this patent application, they are not critical to achieving delayed haloform release.

[0338] At bromoform concentrations in beeswax below 50% by weight, less bromoform was absorbed, suggesting that the mobility of bromoform is limited at low concentrations in beeswax and that beeswax has a strong retention capacity for bromoform (Figure 16B). As the bromoform concentration in beeswax increased and the mass fractions of PBAT and PBS in PLA increased, the mass of bromoform absorbed increased, as did the maximum absorption rate (Figures 16C and D). The absorbed mass of the PLA blend was lower than that of 2003D PLA and 3D printed PLA (Figure 16E).

[0339] Example 5 method Unless otherwise indicated, samples were prepared and analyzed as described in Example 4.

[0340] The hardness of the sample was tested using a Shore D hardness tester at a weight of 7 kg, and structural properties were also tested using XRD before and after exposure to the bromoform / beeswax mixture.

[0341] For XRD analysis, a PANalytica Empyrean XRD with a flat sample stage holder and an adjustable beam to maintain a 1cm x 5mm exposure area at all angles between 5 and 70 2Theta was used in the following configuration: [Table 13]

[0342] The XRD data was exported to Excel, smoothed with 10-point smoothing, and the baseline was corrected between 5 and 60 2theta. result Figure 17 shows the hardness analysis of the PLA blend before and after exposure to bromoform. Therefore, including PBS in the support may reduce the mixture's susceptibility to exposure to bromoform and potentially accelerate its shelf life.

[0343] Example 6: Enhanced high-volume bolus emission test (Rissington test) The bolus was drawn in Solidworks, converted to an .stl file, opened in FlashPrint, and a print job was created. The bolus was printed using 100% filled E-Sun PLA+ at standard resolution, with a first layer height of 0.27mm, layer height of 0.18mm, two outer shell layers, three upper solid layers, three lower solid layers, a hexagonal filling pattern, a print speed of 60mm / s, an extruder temperature of 200°C, and a plate temperature of 50°C, in three separate parts (case, internal structure, and cap).

[0344] Two separate formulations were prepared containing 67% and 55% (by weight) bromoform in a castor wax:paraffin wax mixture (in this example, the ratio was 50:50) as a carrier mixture. Next, after inserting a zinc rod as a densifying agent, the individual bromoform wax mixtures were injected into 1 mm thick casings. The casings were sealed by attaching caps using a soldering gun. The release test was carried out using the method described in Example 1 with some modifications, but using 2 L of medium instead, which was changed daily. A 10 ml sample was taken, properly extracted with ethyl acetate, and then injected into a GC to quantify the bromoform release.

[0345] Bromoform was released at a higher rate from a bolus of 67% (by weight) bromoform (1150 mg / day). On the other hand, the release rate slowed down from a bolus of 55% (by weight) bromoform (9.5 mg / day) (Figure 18A).

[0346] Next, four different types of bolus (two of each) containing 57% (by weight) bromoform with 1 mm and 2 mm casings, and 55% (by weight) and 67% (by weight) bromoform with 2 mm casings were administered using zinc rods with similar carrier compositions. Release tests were performed according to the method described above. The 2 mm casing showed a slow release rate, and it was found that the release rate was slow regardless of the bromoform content (Figures 18B-D). On the other hand, the 1 mm casing bolus containing 57% (by weight) bromoform had a 7-day delay, reaching 240 mg on day 8 and 400 mg on day 9 (Figure 18D). The cumulative plots for days 7, 8, and 9 showed the best fit, exhibiting a release rate of 319 mg / day (Figure 18C).

[0347] Each bolus was tested in RME according to the method described in the above examples. After a 6-day study, the boluses were retrieved and visually inspected. The boluses remained intact with no signs of damage or deformation. Example 7: Design of bromoform-containing boluses In one preferred embodiment tested in this example, the bolus comprises a housing and core configured as defined below. [Table 14]

Claims

1. A delayed-release dosage form or bolus configured for administration to an animal, wherein the dosage form and the bolus are configured to release a hydrophobic substance to the animal over a period of time.

2. A delayed-release dosage form or bolus for administration to ruminants, wherein the delayed-release dosage form and the bolus are configured to release an effective amount of the substance.

3. A delayed-release dosage form or bolus for administration to ruminants, The delayed-release dosage form and the bolus are: A core containing at least one substance to be administered to a ruminant animal mixed with a carrier; and Includes a housing that covers at least part of the core, A bolus is a delayed-release formulation or bolus configured to release a substance through a housing over a predetermined period of time.

4. A bolus according to any one of claims 1 to 3, wherein the carrier and the material have a relatively high affinity for each other compared to the affinity between the housing and the material.

5. The bolus according to claim 4, wherein the substance is a hydrophobic substance.

6. The bolus according to any one of claims 1 to 5, wherein the substance is at least one inhibitor.

7. The bolus according to claim 6, wherein the inhibitor is a haloform, which is preferably selected from the list of bromoform, chloroform, iodoform, and combinations thereof.

8. The bolus according to claim 7, wherein at least one inhibitor is bromoform.

9. The bolus according to claim 7 or 8, wherein a haloform, preferably bromoform, is contained in the core in an amount of 30% to 80% by weight, preferably 30% to 70% by weight.

10. A bolus according to any one of claims 7 to 9, wherein the core contains a haloform, preferably bromoform, in an amount of up to 55% by weight.

11. A bolus according to any one of claims 7 to 10, wherein a haloform, preferably bromoform, is contained in the core, and the carrier contains or consists of wax.

12. The bolus according to any one of claims 3 to 11, wherein the carrier is a polar substance.

13. The bolus according to claim 12, wherein the support comprises a polar functional group such as an ester, an alcohol, or a carbonyl group.

14. The bolus according to any one of claims 3 to 13, wherein the carrier is selected from the group consisting of myristic acid, stearic acid, steryl alcohol, cetyl alcohol, cetosteryl alcohol, castor wax, beeswax, paraffin wax, PEG4000, carnauba, candelilla, jojoba, lanolin, and combinations thereof.

15. The bolus according to claim 14, wherein the wax is mixed with a haloform (preferably bromoform), and preferably the carrier comprises beeswax, paraffin wax and / or castor wax, and more preferably the carrier comprises castor wax and paraffin wax in a weight ratio of castor wax to paraffin wax between 40:60 and 60:

40.

16. The bolus according to claim 11 or 12, wherein the carrier comprises paraffin wax and castor wax.

17. A bolus according to any one of claims 13 to 16, wherein the bolus is at least one metal piece.

18. The bolus according to any one of claims 3 to 17, wherein the housing includes a cavity in which at least a portion of the core is disposed.

19. A bolus according to any one of claims 3 to 18, wherein the housing includes an open end.

20. A bolus according to any one of claims 1 to 19, wherein the bolus includes a cap configured to close the open end.

21. The bolus according to any one of claims 1 to 20, wherein the housing and cap substantially or completely cover and surround the core, thereby defining the core.

22. The bolus according to any one of claims 3 to 21, wherein the housing completely covers and surrounds the core.

23. The bolus according to any one of claims 3 to 22, wherein the housing is formed from a material having a Shore D hardness of at least 40.

24. The bolus according to any one of claims 3 to 23, wherein the housing is formed from a material having a Shore D hardness of less than 70.

25. The bolus according to any one of claims 3 to 24, wherein the housing is formed from a material on which the inhibitor can move.

26. The bolus according to any one of claims 3 to 25, wherein the housing is made of a plastic material.

27. The bolus according to claim 26, wherein the plastic is one or more of polylactic acid (PLA), polyglycolic acid (PGA), polyglycolic acid polylactic acid (PLGA), polypropylene, SLA polymer, PBS, PBAT, or a combination thereof.

28. The bolus according to any one of claims 3 to 27, wherein the housing is made from a material comprising polylactic acid (PLA) and polybutylene adipate terephthalate (PBAT), and the carrier comprises or consists of wax.

29. The bolus according to claim 28, wherein the material contains polylactic acid (PLA) and polybutylene adipate terephthalate (PBAT) in a polylactic acid (PLA):polybutylene adipate terephthalate (PBAT) ratio in the range of 100:0 to 40:60 by weight, and the carrier contains wax.

30. The bolus according to any one of claims 3 to 29, wherein the housing is made from a material containing one or more excipients.

31. The bolus according to claim 30, wherein one or more excipients include a plasticizer, a curing agent and / or a coloring agent.

32. The bolus according to any one of claims 3 to 31, wherein the housing has a material thickness of less than 2 mm, preferably in the range of 0.3 to 1.5 mm.

33. The bolus according to any one of claims 3 to 32, wherein the housing is configured to be disassembled over a predetermined period of time.

34. A bolus according to any one of claims 3 to 33, wherein the core has a melting point above 37°C.

35. The bolus according to any one of claims 3 to 34, further comprising a barrier layer between at least a portion of the housing and the core to isolate the portion of the housing from contact with each other.

36. The bolus according to any one of claims 7 to 35, wherein the bolus is adapted so that the maximum release rate of bromoform into the lumen reaches about 0.05 g to 2 g per day.

37. The bolus according to claim 36, wherein the bolus is adapted to reach a maximum release rate of about 0.1 to 0.5 g of bromoform per day into the lumen, preferably about 0.2 to 0.3 g of bromoform per day into the lumen.

38. A bolus according to any one of claims 1 to 37, wherein the bolus is adapted to release a substance over a period of at least two months.

39. The delayed-release formulation according to any one of claims 1 to 3, wherein the substance is a substance defined in any one of claims 4 to 38 (most preferably bromoform), the core is a core defined in any one of claims 4 to 38, and the housing is a housing defined in any one of claims 4 to 38.

40. A method for administering a substance to an animal, comprising the step of administering to the animal a bolus according to any one of claims 1 to 38, or a delayed-release dosage form according to any one of claims 1 to 3 or 39.

41. A method for reducing methane production in ruminants, The method comprising the step of administering to a ruminant animal a bolus according to any one of claims 1 to 39, or a delayed-release dosage form according to any one of claims 1 to 3 or 39.

42. Use of methane inhibitors and carriers in boluses to reduce methane production in ruminants.

43. Use of methane inhibitors and carriers in boluses to reduce methane emissions from ruminants.

44. The use of haloform in the manufacture of boluses to reduce the emission of one or more greenhouse gases ("GHGs") from ruminants.

45. A method for manufacturing a bolus according to any one of claims 1 to 38, a. To form a housing having a cavity, b. Forming a core containing material, c. Transferring the core to the cavity. The method, including the method described above.

46. The method according to claim 45, wherein the step of forming a core includes mixing a carrier material with a substance.

47. The method according to claim 45 or 46, wherein the step of forming the core includes heating the carrier material to melt it before mixing the carrier material with the substance to produce a mixture.

48. The method according to any one of claims 45 to 47, wherein the step of transferring the core to the cavity includes injecting the mixture into the cavity.

49. A delayed-release dosage form adapted for administration to ruminants, wherein the system comprises a mixture of wax and haloform.

50. A delayed-release dosage form adapted for administration to a ruminant according to claim 49, the delayed-release dosage form comprising: a core comprising a wax and a haloform (preferably bromoform); and a coating covering at least a portion of the core, preferably covering the entire core; the delayed-release dosage form releasing a haloform.

51. The delayed-release agent type according to claim 50, wherein the coating is the housing according to any one of claims 4 to 38, and the core is the core according to any one of claims 4 to 38.

52. The delayed-release formulation according to any one of claims 49 to 51, wherein the haloform is bromoform.

53. A delayed-release dosage form according to any one of claims 49 to 52, or a bolus according to any one of claims 3 to 38, The carrier contains or consists of wax, and the coating / housing contains PLA, PBAT, or a mixture of both; preferably, the coating / housing contains PLA, the delayed-release formulation or bolus.

54. A delayed-release dosage form according to any one of claims 1, 2, 39, 49 to 53, or a bolus according to any one of claims 1 to 38, A delayed-release formulation or bolus wherein the core of the delayed-release formulation or bolus contains one or more metal particles (preferably steel particles), the particles are preferably round, and the total mass of all particles per bolus or per delayed-release formulation is at least 100 grams.

55. The delayed-release formulation according to claim 54, wherein the particles are granules and / or spheres.

56. The delayed-release formulation according to any one of claims 49 to 55, wherein the wax is paraffin and / or carnauba and / or castor wax.

57. The delayed-release formulation according to any one of claims 49 to 56, wherein the core comprises 30% to 75% by weight of haloform, preferably bromoform.

58. The delayed-release formulation according to any one of claims 49 to 57, wherein the thickness of the coating / housing is less than 2 mm.

59. The delayed-release dosage form according to any one of claims 49 to 58, wherein the delayed-release dosage form has a bolus shape.

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