Improved bolus design and its use for administration to ruminants

Abstract

This disclosure provides an improved bolus configured for administration to animals, wherein the bolus is configured to release a methane inhibitor or another active agent to the animal. Preferably, the methane inhibitor is a haloform, but the bolus may be adapted as described herein to accommodate other active agents, such as other methane inhibitors. Methods for using and producing the bolus are also provided.

Description

[Technical Field]

[0001] This disclosure relates to improvements in devices and methods for delivering substances to animals, specifically to devices and methods for administering at least one substance to the rumen of a ruminant, as well as to methods for manufacturing such devices. [Background technology]

[0002] In livestock farming, it is often necessary to deliver substances to animals. This may be for any of a variety of purposes, including but not limited to the treatment or prevention of disease, and to increase animal productivity.

[0003] There are various devices (e.g., dosage forms) and methods for delivering substances such as drugs to animals. Some substances are for administration to the rumen of ruminants. Some dosage forms are for sustained release (i.e., continuous release) to the rumen of ruminants. Some dosage forms are for the administration of multiple active ingredients to the rumen of ruminants. However, there is still a need for the administration of substances to animals without requiring repeated administration by farmers or animal husbands. Sustained-release dosage forms present challenges regarding extended control of the released dose, as well as the reliability of such control over time and between batches. Similarly, controlling the release of multiple active ingredients also presents challenges in terms of release profiles and reliability.

[0004] The sustained release and / or release of multiple active ingredients within the rumen presents further challenges, particularly for sustained-release formulations, due to the less-studied local environment compared to the gastrointestinal tract of monogastric organisms (e.g., humans). Controlled sustained release of substances within the rumen can increase the potency of the administered formulation and reduce side effects by reducing the maximum concentration of the active ingredient to a concentration more consistent with the effective concentration and maintaining such an effective concentration over an extended period. The release of multiple active ingredients from a single formulation reduces the number of doses required. Administering formulations to larger animals, such as ruminants, may require significant investment in time and infrastructure. However, the possibility that multiple active ingredients may influence the release profile complicates such formulations, especially in sustained-release formulations.

[0005] There is a need for improved dosage forms for the sustained release and / or release of multiple active ingredients into the lumen. Preferably, these dosage forms provide controlled sustained release into the lumen, and more preferably, the control is maintained until the majority of the substance has been released from the dosage form into the lumen.

[0006] In some cases, sustained release of a low dose of the active substance over an extended period may be particularly desirable, as this can provide the best efficacy of the treatment.

[0007] The challenges associated with sustained release of a substance increase with the desired extension. Extending release over hours, days, weeks, and months becomes increasingly difficult, as does controlling the release to deliver a biologically effective and non-toxic dose over the extended timeframe. There is a need for improved dosage forms for delivering substances with a sustained-release profile to the rumen of ruminants over extended periods. Preferably, the sustained release is well controlled over that period.

[0008] No prior art reference in this specification constitutes an endorsement or suggestion that the prior art forms part of common sense in any jurisdiction, or that the prior art can be reasonably expected to be understood, considered relevant, and / or combined with other prior art by those skilled in the art.

[0009] One aspect of the present invention is to provide improved devices and methods for delivering substances, such as methane inhibitors, to animals.

[0010] One aspect of the present invention is to provide devices and methods for reducing greenhouse gas ("GHG") emissions.

[0011] For example, one aspect of the present invention is to provide devices and methods for improving animal production gains through the reduction of methane production.

[0012] One aspect of the present invention is to provide a formulation for reducing GHG excretion by one or more animals, such as ruminants.

[0013] One aspect of the present invention is to provide a device and method capable of releasing a substance at different rates over a certain period of time, such as a sustained-release formulation containing a methane inhibitor.

[0014] Providing devices and methods adapted to the specific properties of certain GHG inhibitors is also an aspect of the present invention.

[0015] Alternatively, one aspect of the present invention is to overcome some of the shortcomings of the prior art.

[0016] Alternatively, one aspect of the present invention is to provide the public with useful options for methane inhibitors, as well as suitable devices and methods for administering these methane inhibitors. [Overview of the project]

[0017] This disclosure relates to a device and method for delivering a substance to an animal. In a preferred form further outlined herein, the substance is a methane inhibitor. This disclosure is illustrated by reference to preferred embodiments; however, these should not be seen as limitations on the scope of this disclosure. All documents referenced herein are incorporated by reference. Unless otherwise claimed, all embodiments disclosed herein can be combined.

[0018] In a first aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core containing a methane inhibitor; and a housing surrounding (or covering or completely enclosing) the core, wherein the housing is configured to increase its permeability to the methane inhibitor when the housing is exposed to the rumen of a living animal; and / or, wherein at least a portion of the housing is configured to form one or more openings when the housing is exposed to the rumen of a living animal, allowing an increased release of the methane inhibitor from within the bolus through the opening(s).

[0019] In a further aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: A core containing a methane inhibitor; and a housing covering at least a portion of the core. Here, the housing is configured such that, when exposed to a phosphate buffer (pH: 6.5, 0.02 M) at 40°C without stirring, its permeability to the methane inhibitor increases by at least 5% relative to its permeability on the same day after exposure to the phosphate buffer (pH: 6.5, 0.02 M) at a reference temperature without stirring for the same duration. Here, permeability is evaluated based on the release rate of the methane inhibitor.

[0020] In a further aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, and wherein the bolus comprises: a core, wherein the core comprises the methane inhibitor in microencapsulated particles, the microencapsulated particles are dispersed in a composition comprising a carrier and optionally also comprising a dispersant; and a housing covering at least a portion of the core.

[0021] In a further aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, and wherein the bolus comprises: a carrier and microencapsulated particles, wherein the microencapsulated particles comprise the methane inhibitor, the microencapsulated particles are dispersed in the carrier, and preferably, the carrier has a porous structure such as mesoporous silica.

[0022] In a further aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises a core, the core comprises the methane inhibitor and a carrier, and the bolus does not comprise a housing.

[0023] In a further aspect, the present disclosure relates to a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, and wherein the bolus comprises: a core, wherein the core comprises the methane inhibitor and a carrier; and a housing containing the core; wherein the methane inhibitor is selected from the group consisting of monensin, lauric acid, myristic acid, and linoleic acid; wherein the housing comprises at least one opening that exposes the core to the environment surrounding the bolus.

[0024] In a further aspect, the present disclosure relates to the bolus of the present disclosure, wherein in addition to or instead of said methane inhibitor, the bolus comprises an active agent, wherein said active agent is selected from the group consisting of anti-inflammatory agents, analgesics, antibiotics, and anthelmintics.

[0025] In a further aspect, there is provided a bolus for administration to a ruminant, wherein said bolus is configured to release a methane inhibitor in an animal, wherein said bolus comprises: a core; a housing covering at least a portion of the core: wherein the core comprises at least one methane inhibitor and at least one further active agent.

[0026] In a further aspect, there is provided a bolus for administration to a ruminant, wherein said bolus is configured to release a methane inhibitor in an animal, wherein said bolus comprises: a core comprising a methane inhibitor dispersed in, or forming part of, one or more of a hydrogel, oleogel, or organogel; and a housing covering at least a portion of the core.

[0027] In another aspect, the present disclosure relates to the bolus of the present disclosure for use in the treatment of an animal, preferably a ruminant. In another aspect, the present disclosure relates to the bolus of the present disclosure for use in reducing and / or inhibiting methane emission in ruminants. The present disclosure also provides a method of treating an animal comprising administering the bolus of the present disclosure to said animal.

[0028] In a further aspect, the present disclosure relates to a method of manufacturing the bolus of the present disclosure.

[0029] In a further aspect, the present disclosure relates to methane inhibitors for use in reducing methane emissions from ruminants, wherein the methane inhibitor is selected from the group consisting of bromoform, monensin, lecithin, lauric acid, myristic acid, linoleic acid, and phospholipids.

[0030] In a further aspect, the disclosure relates to a method for treating ruminants to reduce methane emissions from said ruminants, comprising administering to said animals a methane inhibitor selected from the group consisting of bromoform, monensin, lecithin, lauric acid, myristic acid, linoleic acid, and phospholipids.

[0031] In another aspect, the Disclosure provides a method for administering a methane inhibitor to a ruminant, the method comprising administering a bolus in accordance with the Disclosure to the rumen of the ruminant.

[0032] In another aspect, the disclosure provides a method for reducing methane production in the rumen of a ruminant, the method comprising administering a bolus in accordance with the disclosure to the rumen of a ruminant.

[0033] In yet another aspect, this disclosure provides a method for making a bolus, the method comprising: Selecting the core and housing, Inserting the core into the housing, Optionally, sealing the housing to enclose or substantially enclose the core inside the housing: Here, the core contains a methane inhibitor. Optionally, the core is melted and poured into a housing, where it solidifies. Optionally, the housing surrounds the core after it has solidified.

[0034] In some embodiments, the housing is configured to increase its permeability to the methane inhibitor when the housing is exposed to the lumen of a living animal.

[0035] In a further aspect, materials comprising acrylate polymers are provided for use in packing the boluses of the present disclosure.

[0036] Further aspects of the present disclosure described in the preceding paragraphs and further embodiments thereof will become apparent from the following description, given by example and with reference to the accompanying drawings. [Brief explanation of the drawing]

[0037] [Figure 1] Release profiles of boluses loaded with a matrix composed of 60% tribromomethane, 20% EC, and 20% HPMC, when placed in a buffer at room temperature (RT: 25°C), 30°C, and 40°C. [Figure 2] Release profiles of boluses loaded with a matrix composed of 58.4% tribromomethane, 27.3% HPMC, and 14.3% EC, when placed in a buffer at room temperature (RT: 25°C), 30°C, and 40°C. [Figure 3] Emission profiles of boluses with housing thicknesses of 0.9, 1.2, and 1.5 mm loaded with a matrix composed of 58.4% tribromomethane, 27.3% HPMC, and 14.3% EC, when placed in a buffer at 40°C. [Figure 4] An inverted vial containing lecithin-TBM gel. [Figure 5] Release profiles of boluses loaded with matrices composed of 55% lecithin / 45% tribromomethane, 40% lecithin / 60% tribromomethane, and 30% lecithin / 70% tribromomethane over 30 days. [Figure 6] Release profile of a bolus loaded with a matrix composed of 55% lecithin and 45% tribromomethane over 120 days. [Figure 7] An inverted vial containing PMMA-TBM gel. [Figure 8]Release profiles of boluses loaded with matrices composed of 55% PMMA / 45% tribromomethane and 30% PMMA / 70% tribromomethane over 45 days. [Figure 9] Release profile of a bolus loaded with a matrix composed of 55% PMMA / 45% tribromomethane over 90 days. [Figure 10] Release profiles of boluses loaded with matrices composed of 30% MCW / 70% tribromomethane and 35% MCW / 65% tribromomethane over 30 days. [Figure 11] Release profile of a bolus loaded with a matrix composed of 35% MCW / 65% tribromomethane over 80 days. [Figure 12] Release profiles of boluses loaded with 17, 34, 52, and 70 g MCW 35% / tribromomethane 65% matrices. [Figure 13] Release profiles of boluses loaded with matrices composed of 30% stearic acid / 70% tribromomethane and 35% stearic acid / 65% tribromomethane. [Figure 14] Changes in the release profile of a bolus loaded with 35% eicosane and 65% tribromomethane when the temperature is raised from room temperature to 40°C. [Figure 15] Emission profiles for MCW 35% / tribromomethane 65% with or without housing (PLA / PBAT=9 / 1). [Figure 16] Emission profiles for MCW 55% / tribromomethane 45% with or without housing (PLA / PBAT=9 / 1). [Figure 17] Effect of lecithin (5% w / w) inclusion on the release profile of a stearic acid matrix loaded with 65% TBM. The increase in lecithin concentration (5%) was compensated for by a corresponding decrease in stearic acid from 35% to 30% w / w. [Figure 18]The effect of monensin (2% and 5% w / w) inclusion on the release profile of TBM-loaded EC / HPMC matrices. Increases in monensin concentration (0, 2, and 5%) were compensated for by corresponding decreases in HPMC content (20, 18, and 15%). EC and TBM concentrations remained constant at 20% and 60% w / w, respectively. [Figure 19] The effect of inclusion of phloroglucinol (2% and 5% w / w) on the release profile of TBM-loaded EC / HPMC matrices. Increases in phloroglucinol concentration (0, 2, and 5%) were compensated for by corresponding decreases in HPMC content (20, 18, and 15%). EC and TBM concentrations remained constant at 20% and 60% w / w, respectively. [Figure 20] The effect of albendazole (2% and 5% w / w) inclusion on the release profile of TBM-loaded EC / HPMC matrices. Increases in albendazole concentration (0%, 2%, and 5%) were compensated for by corresponding decreases in HPMC content (20%, 18%, and 15%). EC and TBM concentrations remained unchanged at 20% and 60% w / w, respectively. [Figure 21] The effect of ketoprofen (2% and 5% w / w) inclusion on the release profile of TBM-loaded EC / HPMC matrices. Increases in ketoprofen concentration (0, 2, and 5%) were compensated for by corresponding decreases in HPMC content (20, 18, and 15%). EC and TBM concentrations remained unchanged at 20% and 60% w / w, respectively. [Figure 22] A bolus (100) of the present disclosure comprising a housing (101), a core (102), a sealed region (103), and a densifying agent (104). [Figure 23] Emission profiles from boluses with high amorphous or high crystalline PLA content casings loaded with 60g of 65% TBM / 35% MCW over 12 days. [Modes for carrying out the invention]

[0038] Certain substances present particular difficulties in the context of sustained release to the rumen. One class of compounds that are difficult to deliver to animals is hydrophobic compounds. A further class of compounds that are particularly difficult to deliver to animals in sustained release is volatile or partially volatile compounds. The properties of these compounds present challenges for developing techniques for the sustained release of these hydrophobic and / or volatile / partially volatile substances, particularly through the stomach of animals. Haloforms such as bromoform are such substances. Surprisingly, the inventors have developed extremely long-lasting sustained-release (several months) dosage forms for delivering hydrophobic and / or volatile / partially volatile substances to the rumen of ruminants, an environment that has been relatively little studied compared to the gastrointestinal tract of monogastric animals. Surprisingly, the inventors have improved the release kinetics and duration of controlled release of the dosage form through the development of core components.

[0039] One specific purpose of administering substances to animals is to reduce the harmful effects of agriculture. For example, it is well known that various methane and nitrification inhibitors are administered to animals to reduce or mitigate the harmful effects of methane and nitrogen-containing compounds produced by them.

[0040] However, despite current efforts, climate change is creating widespread environmental and social impacts on a global scale. It is widely understood that these impacts will only continue to increase over time. Consequently, there is a global push to reduce harmful greenhouse gas (GHG) emissions in efforts to mitigate the worst effects of climate change.

[0041] The agricultural sector is considered a major source of GHG emissions. Global methane emissions from livestock are estimated to contribute 7.1 gigatons of CO2 equivalent per year, representing 14.5% of all anthropogenic greenhouse GHG emissions. Therefore, this sector will play a crucial role in reducing overall GHG emissions.

[0042] The main GHGs released by agriculture are methane (CH4) and nitrous oxide (N2O), with livestock being the primary source of methane emissions. Most methane is released when cattle or other ruminants burp. The amount of methane produced for each farm is directly related to total animal feed intake, which is usually measured as dry matter intake (DMI).

[0043] Countries with strong agricultural sectors, such as New Zealand and others, face challenging goals in reducing agricultural emissions. For example, the New Zealand government has implemented policies aiming to reduce methane emissions by 24-50% before 2050. In New Zealand, too, livestock methane production is estimated to account for about half of the country's total GHG emissions. Reducing methane is a critical component in meeting GHG emission targets and mitigating the effects of global warming.

[0044] The release of GHGs by animals also has detrimental effects on animal productivity. Any feed that is subsequently converted into compounds exhaled or released by animals is an energy source that has not been converted into productive use. Therefore, for efficiency, it is important to optimize the conversion of feed into animal productivity, including in the form of weight gain or milk production.

[0045] Conventional devices for administering methane inhibitors or other active ingredients to animals, known from the literature, can still be further improved in terms of, for example, durability, control of the rate of release of the active agent, versatility of administering drug forms such as boluses, and reduction of size and manufacturing costs for making draft forms or, preferably, boluses. definition

[0046] Unless otherwise defined herein, the following terms shall be understood to have the following general meanings:

[0047] As used herein, “carboxylate glass” means glass formed when one or a mixture of metal carboxylates is heated above their melting point and allowed to cool.

[0048] As used herein, "bromoform-rich seaweed extract" refers to an extract containing non-trace seaweed components other than bromoform. Bromoform with a purity of at least 90% (w / w), at least 95% (w / w), at least 96% (w / w), and at least 99% (w / w) is not considered a "bromoform-rich seaweed extract."

[0049] An "active agent" may be any substance that provides a benefit to an animal, for example, a drug for treating or preventing a disease that improves animal productivity or mitigates at least one harmful effect of agriculture. For example, an active agent may modulate the metabolism of an animal and affect, for example, the quantity or quality of methane production.

[0050] Veterinary-acceptable excipients are those that, when administered to an animal subject, are typically not harmful to the subject. Those skilled in the art will understand that veterinary-acceptable excipients generally include pharmaceutically acceptable (i.e., human-acceptable) excipients.

[0051] As used herein, “haloform” is CHX3, where X is a halogen, and each X atom may be a different halogen. Therefore, “haloform” encompasses CHClBr2 and similar compounds. As used herein, “mixed haloform” means a haloform in which not all X atoms bonded to the carbon atoms are the same. In some embodiments, each X atom is the same. The terms “bromoform” and “tribromomethane” are used interchangeably herein.

[0052] As used herein, “to break down” and “to decompose” do not require the complete breakdown of the bolus into other substances that are completely absorbed by the rumen fluid, but rather require only that the bolus be sufficiently broken down so that it can leave the rumen, for example, by passing through or refluxing the digestive tract of an animal.

[0053] As used herein, “feed” means dry matter intake (DMI), supplements, pasture grass, grains, or other feed stocks.

[0054] As used herein, the term “effective dose” means the amount of an active ingredient that would induce a biological or medical response in a tissue, system, or animal, as sought, for example, by a researcher or veterinarian. Furthermore, the term “therapeutic dose” means any amount that results in an improved treatment, cure, prevention, or improvement of a disease, disorder, or side effect, or a reduction in the rate of progression of the disease or disorder, compared to a corresponding subject that has not received such a dose. The term also encompasses amounts that are effective in enhancing normal physiological function. For example, a therapeutic dose of a methane inhibitor such as haloform reduces methane production in animals, preferably ruminants.

[0055] It should also be noted that the singular forms “a,” “an,” and “the” as used herein and in the appended claims encompass multiple references unless the context clearly specifies otherwise. For example, the reference to “polymer” may encompass multiple polymers, and the reference to “at least one carrier” may encompass one or more carriers.

[0056] The term "and / or" can mean "and" or "or".

[0057] The term "(singular or plural)" following a noun intends for it to be singular, plural, or both.

[0058] As used herein, “methane inhibitor” refers to an active agent, such as a compound or mixture of compounds, that can inhibit or reduce the production of methane gas in the rumen of ruminants. Methane inhibitors can suppress methane formation. As used herein, “methane inhibitor” is preferably a haloform, more preferably a bromoform.

[0059] The term "enveloping the core" can mean completely enclosing the core. This disclosure also intends to describe partially enclosing the core.

[0060] Except where the context otherwise requires, the terms “comprise” and variations thereof, such as “comprising,” “comprises,” and “comprised,” as used herein are not intended to exclude further additives, components, integers, or steps.

[0061] Various features of this disclosure are described by reference to certain values ​​or ranges of values. These values ​​are intended to relate to the results of various appropriate measurement techniques and should therefore be interpreted as encompassing the tolerances inherent in any particular measurement technique. Some of the values ​​referenced herein are expressed by the term “about” to at least partially describe this variability. When used to describe a value, the term “about” may mean a quantity within ±25%, ±10%, ±5%, ±1%, or ±0.1% of that value. Increased permeability and / or aperture

[0062] In a first aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core containing a methane inhibitor; and a housing surrounding (or covering or completely enclosing) the core, wherein the housing is configured to increase its permeability to the methane inhibitor when the housing is exposed to the rumen of a living animal; and / or, wherein at least a portion of the housing is configured to form one or more openings when the housing is exposed to the rumen of a living animal, allowing an increased release of the methane inhibitor from within the bolus through the opening(s).

[0063] In a further aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: A core containing a methane inhibitor; and a housing covering at least a portion of the core. Here, the housing is configured such that, when exposed to a phosphate buffer (pH: 6.5, 0.02 M) at 40°C without stirring, its permeability to the methane inhibitor increases by at least 5% relative to its permeability on the same day after exposure to the phosphate buffer (pH: 6.5, 0.02 M) at a reference temperature without stirring for the same duration. Here, permeability is evaluated based on the release rate of the methane inhibitor.

[0064] In some embodiments, the reference temperature is 20°C, 25°C, or 30°C. In some embodiments, the difference in permeability of the housing is evaluated based on the emission rate for day 7, day 14, day 30, or day 60. In some embodiments, the difference in permeability is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%.

[0065] A bolus for administration to ruminants is further provided, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core containing the methane inhibitor; and a housing surrounding (or covering at least some portion thereof or completely enclosing) the core, wherein the housing is configured to increase its permeability to the methane inhibitor when exposed to a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C; and / or, wherein at least a portion of the housing is configured to form one or more openings when exposed to a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C, allowing the methane inhibitor to exit the bolus through the opening(s).

[0066] In some embodiments, the housing surrounds the core.

[0067] The release behavior of the active agent released from the bolus to the outside of the bolus can be determined by submerging a bolus according to this disclosure, containing the active agent, in a tank filled with 25 liters of phosphate buffer (pH: 6.5, 0.02 M) at a constant temperature of 40°C, where the liquid buffer surrounding the bolus can be continuously stirred using a magnetic stirrer. After a given time, the concentration of the active agent in the phosphate buffer is quantified (e.g., using GC-FID (gas chromatography connected to a flame ionization detector)). The quantification can be repeated at certain intervals, for example, once daily.

[0068] In a preferred embodiment, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core containing the methane inhibitor; and a housing surrounding (or covering or completely enclosing) the core, wherein the housing is configured to increase its permeability to the methane inhibitor when the housing is exposed to a temperature of at least 38°C; and / or, wherein at least a portion of the housing is configured to form one or more openings when the housing is exposed to a temperature of at least 38°C, allowing the methane inhibitor to exit the bolus through the opening(s).

[0069] Certain embodiments disclosed herein relate to a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core containing the methane inhibitor; and a housing surrounding (or covering or completely enclosing at least a portion thereof) the core, wherein the housing is permeable to the methane inhibitor when the housing is exposed to a temperature of at least physiological or lumen temperature; and / or, wherein at least a portion of the housing is configured to form one or more openings when the housing is exposed to a temperature of at least physiological or lumen temperature, allowing the methane inhibitor to exit the bolus through the opening(s). In a preferred embodiment, the housing surrounds the core.

[0070] In a further aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core containing a methane inhibitor; and a housing surrounding (or covering or completely enclosing) the core, wherein the permeability of the housing to the methane inhibitor increases when the housing is exposed to a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C; and / or, wherein at least a portion of the housing is configured to form one or more openings when the housing is exposed to a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C, allowing for increased release of the methane inhibitor from within the bolus through the opening(s).

[0071] In a preferred embodiment, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, and the bolus comprises: a core containing the methane inhibitor; and a housing surrounding (or completely covering or enclosing at least a portion thereof) the core, wherein the permeability of the housing to the methane inhibitor increases when the housing is exposed to a temperature of at least 38°C; and / or, wherein at least a portion of the housing is configured to form one or more openings when the housing is exposed to a temperature of at least 38°C, allowing increased escape of the methane inhibitor from the bolus through the opening(s).

[0072] In the preferred embodiment of the bolus type described above, the housing includes a plurality of openings, each having an average diameter between, for example, 1 micrometer and 0.5 mm, where each opening is filled with a substance that melts at a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C, such as wax or a hydrocarbon composition. In an alternative or additional preferred embodiment of the bolus, the housing including the openings is surrounded by a film of wax or hydrocarbons, such as a wax or hydrocarbon composition, which melts at a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C. Increased permeability

[0073] In some embodiments, the permeability of the housing to the methane inhibitor increases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% relative to the permeability on the same day after exposure of the housing to a phosphate buffer (pH: 6.5, 0.02 M) at 20°C without agitation for the same length of time when the housing is exposed to the rumen of a living animal, Here, permeability is evaluated based on the release rate of the methane inhibitor. In some embodiments, the permeability of the housing to the methane inhibitor increases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% relative to the permeability on the same day after exposure of the housing to a phosphate buffer (pH: 6.5, 0.02 M) at 25°C without agitation for the same length of time when the housing is exposed to the rumen of a living animal, Here, permeability is evaluated based on the release rate of the methane inhibitor.

[0074] In some embodiments, the permeability of the housing to the methane inhibitor increases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% relative to the permeability on the same day after exposure of the housing to a phosphate buffer (pH: 6.5, 0.02 M) at 28°C without agitation for the same length of time when the housing is exposed to the rumen of a living animal, Here, permeability is evaluated based on the release rate of the methane inhibitor.

[0075] In some embodiments, the permeability of the housing to the methane inhibitor increases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% relative to the permeability on the same day after exposure of the housing to a phosphate buffer (pH: 6.5, 0.02 M) at 30°C without agitation for the same length of time when the housing is exposed to the rumen of a living animal, Here, permeability is evaluated based on the release rate of the methane inhibitor. In some embodiments, the permeability of the housing to the methane inhibitor increases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% relative to the permeability on the same day after exposure of the housing to a phosphate buffer (pH: 6.5, 0.02 M) at 38°C without agitation for the same length of time when the housing is exposed to the rumen of a living animal, Here, permeability is evaluated based on the release rate of the methane inhibitor.

[0076] In some embodiments, differences in housing permeability are evaluated based on the release rate on day 7, day 14, day 30, or day 60, relative to the release rate on the same day after exposure of the bolus to a phosphate buffer (pH: 6.5, 0.02 M) at a specified temperature without agitation for the same length of time, following exposure of the bolus to the lumens of living animals.

[0077] In some embodiments, the permeability of the housing to the methane inhibitor increases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% relative to the permeability on the same day after exposure to a bolus of phosphate buffer (pH: 6.5, 0.02M) at 20°C without stirring for the same duration when the bolus is exposed to a bolus of phosphate buffer (pH: 6.5, 0.02M) at 40°C without stirring.

[0078] In some embodiments, the permeability of the housing to the methane inhibitor increases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% relative to the permeability on the same day after exposure to a bolus of phosphate buffer (pH: 6.5, 0.02M) at 25°C without stirring for the same duration when the bolus is exposed to a bolus of phosphate buffer (pH: 6.5, 0.02M) at 40°C without stirring.

[0079] In some embodiments, the permeability of the housing to the methane inhibitor increases by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% relative to the permeability on the same day after exposure to a bolus of phosphate buffer (pH: 6.5, 0.02M) at 30°C without stirring for the same length of time when the bolus is exposed to a phosphate buffer (pH: 6.5, 0.02M) at 40°C without stirring.

[0080] In some embodiments, differences in housing permeability are evaluated based on the release rate on day 7, day 14, day 30, or day 60, relative to the permeability on the same day after exposure of a bolus to a phosphate buffer (pH: 6.5, 0.02 M) at a specified temperature for the same length of time without stirring, after exposure of a bolus to a phosphate buffer (pH: 6.5, 0.02 M) at 40°C without stirring for the same length of time.

[0081] The increased permeability of the housing to the methane inhibitor leads to diffusion of the methane inhibitor from the bolus to the outside when the concentration of the methane inhibitor inside the bolus, which is not bound by a carrier, is greater than that outside the bolus. The permeability of the housing to the methane inhibitor can be evaluated by any preferred means, for example, by the diffusion of the methane inhibitor from the bolus to the outside. Diffusion from the bolus to the outside can be measured by any preferred means, for example, by GC-FID analysis of the surrounding mixture. GC-FID is suitable for quantifying bromoform and therefore for quantifying the permeability of the housing to bromoform. Samples from the rumen of living animals can be collected from fistula-wearing animals. housing

[0082] As used herein, the term “housing” is generally understood to mean a casing that surrounds, covers at least a portion of, or completely encloses a core containing at least one methane inhibitor and possibly further active agents. The housing may include a cap.

[0083] In some embodiments, the housing surrounds the core. In some embodiments, the housing substantially surrounds the core. A substantially surrounded core is covered by the housing to about 70–99%, about 80–99%, about 85–99%, or about 90–99% of the core's surface area.

[0084] The housing may be made from a composition including, for example, a biodegradable plastic. In some embodiments, the housing comprises one or more biodegradable polymers. In some embodiments, the housing consists of one or more biodegradable polymers. In some embodiments, the housing does not contain any non-biodegradable polymers. In some embodiments, the housing comprises one or more non-biodegradable polymers comprising polyvinyl chloride (PVC), polyethylene terephthalate (PET), beech S, nylon, polyvinyl butyral, polyethylene (low, medium, high, or ultra-high density), polypropylene (PP), and combinations thereof. In some embodiments, the housing comprises one or more non-biodegradable polymers comprising high-density polyethylene (HDPE), polypropylene (PP), and combinations thereof. Optionally, the non-biodegradable polymers in the housing are combined with biodegradable polymers or are biodegradable in the presence of haloforms and / or cores.

[0085] In some aspects, materials such as plastics are considered biodegradable if they are considered biodegradable under the standards specified in ISO 14855-1:2012 (Biodegradability of plastic materials under controlled composting conditions). According to this method, the percentage of biodegradation is given by the ratio of the CO2 produced from the test material to the maximum theoretical amount of CO2 that can be produced from the test material (excluding the amount of carbon that is converted into new cellular biomass, i.e., not metabolized into CO2). The maximum theoretical amount of CO2 produced is calculated from the total organic carbon content of the test material. The threshold for industrial composting biodegradability is biodegradation of at least 90% by mass of the total mass of the test material in less than 6 months. Therefore, for example, for a test material to be considered biodegradable, 90% of the carbon in the test material should be converted to CO2 within 6 months.

[0086] Preferably, the housing material is compliant with waste disposal regulations applicable to the slaughterhouse. The housing material may generally include any material that is non-toxic when administered to the animal's rumen. Particularly applicable is that any food animal will produce non-toxic food (meat or milk) after exposure to the material in the bolus. The housing material is more preferably thick enough (wall thickness) to resist mechanical stress and abrasive forces in the rumen and to allow it to remain intact and prevent fracturing or collapse inside the rumen for at least several weeks.

[0087] The housing is molded to fit with the core and any other components within the bolus, so that no air pockets exist within the bolus.

[0088] The housing may be made from a material through which a methane inhibitor can move, for example, by a mass diffusion process. In a preferred embodiment, the housing may be made from at least one plastic material, for example, a biodegradable plastic or material that decomposes over time in the lumen. In one embodiment, 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 a combination thereof.

[0089] In any embodiment herein, a bolus comprising a housing described herein may comprise, in or in part thereof, a housing comprising polylactic acid (PLA), polybutylene succinate-co-adipate (PBSA), polybutylene succinate (PBS), polyhydroxybutyrate-co-hydroxyvalerate, polyvinyl acetate (PVA), polybutylene adipate terephthalate (PBAT), polycaprolactone (PCL), wood flour and cellulose materials, ethylcellulose and hydroxypropyl methylcellulose, and at least one compound selected from the group consisting of two or more mixtures of the above.

[0090] In another embodiment, the housing comprises one or more hydrophobic polymers. Optionally, the housing comprises one or more hydrophobic biodegradable polymers. In a different embodiment, the housing consists of one or more hydrophobic polymers. Optionally, the housing consists of one or more hydrophobic biodegradable polymers.

[0091] In some embodiments, the housing comprises one or more ester-based polymers.

[0092] In some embodiments, the housing comprises one or more polymers selected from a list consisting of high-density polyethylene (HDPE), polypropylene (PP), polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), poly-D,L-lactic acid (PDLLA), polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), combinations thereof, and copolymers thereof. In some embodiments, the housing comprises one or more polymers selected from a list consisting of polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), poly-D,L-lactic acid (PDLLA), polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), combinations thereof, and copolymers thereof. In some embodiments, the housing comprises one or more polymers selected from a list consisting of high-density polyethylene (HDPE), polypropylene (PP), combinations thereof, and copolymers thereof.In some embodiments, the housing comprises one or more polymers selected from a list consisting of polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), poly-D,L-lactic acid (PDLLA), polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), combinations thereof, and copolymers thereof. In some embodiments, the housing is made of one or more polymers selected from a list consisting of polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), poly-D,L-lactic acid (PDLLA), polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), combinations thereof, and copolymers thereof.

[0093] In some embodiments, the housing comprises one or more 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 or one or more thermosetting polymers and / or resins, ABS, combinations thereof, and copolymers thereof. In some embodiments, the housing comprises one or more polylactic acid (PLA), polybutylene succinate-co-adipate (PBSA), polybutylene succinate (PBS), polyhydroxybutyrate-co-hydroxyvalerate, polyvinyl acetate (PVA), polybutylene adipate terephthalate (PBAT), polycaprolactone (PCL), wood flour and cellulose materials, ethylcellulose and hydroxypropyl methylcellulose, combinations thereof, and copolymers thereof.

[0094] In some embodiments, the housing comprises one or more polymers selected from the list consisting of polylactic acid, polybutylene adipate terephthalate, combinations thereof, and copolymers thereof.

[0095] In some embodiments, the PLA:PBAT ratio is approximately 95:5 to 70:30 wt / wt, approximately 95:5 to 80:20 wt / wt, or approximately 95:5 to 85:15 wt / wt. In some embodiments, the PLA:PBAT ratio is approximately 90:10 wt / wt.

[0096] Such blends of materials can be particularly advantageous. For example, mixing / blending polybutylene polymers such as PBAT with PLA increases the plasticity and strength of the housing compared to a housing made from PLA alone, while preserving the biodegradability of the housing material. This stability improvement effect is particularly beneficial when using, for example, haloform as a methane suppressant, because such compounds can otherwise promote the brittleness of the housing material. Furthermore, the use of polybutylene polymer / PLA blends improves the durability of the housing compared to PLA alone, reducing the risk of fracture under mechanical stress, for example, when placed in an animal's lumen.

[0097] The components used or mixed to form the housing material may be selected according to their suitability for use in forming the bolus housing. Due to heating for molding the bolus housing, the composition should not become too viscous for 3D printing or injection molding, and blends of two or more polymers should result in a homogeneous mixture without excessive bubbles. 3D printing encompasses stereolithography (SLA) and digital light processing (DLP).

[0098] The bolus housing may include, for example, biodegradable and / or non-biodegradable materials, but preferably a biodegradable polymer. Such materials may be synthetic, natural, or essentially naturally derived. It is preferable that the material be selected from biodegradable polymers. Examples of such polymers include, without limitation, polylactic acid (PLA), polybutylene terephthalate (PBT), polybutylene adipate terephthalate (PBAT), polybutylene succinate (PBS), and polybutylene succinate adipate (PBSA). Biodegradability allows for repeated administration of the bolus while preventing the accumulation of bolus material in the body of the ruminant, as the bolus components may be at least partially or even completely degraded in the rumen environment. Nevertheless, even if the bolus housing is biodegradable, it is understood that it will not be completely degraded when retained in the rumen over a duration of at least 7 days, to the extent that the bolus degrades over this period. Suitable non-biodegradable polymers include high-density polyethylene (HDPE), polypropylene (PP), combinations thereof, and copolymers thereof.

[0099] Accordingly, a housing for any bolus, including the housings described herein, may be configured to have sufficient structural integrity to remain intact over a predetermined period of time. In a preferred embodiment, the housing may be configured to decompose over a predetermined period of time. The predetermined period may mean the period over which the methane inhibitor is to be released to animals. In a particularly preferred embodiment, the predetermined period may be at least two months, preferably six months, and more preferably twelve months. In some embodiments, the period may be at least two weeks, three weeks, four weeks, or six weeks.

[0100] In one embodiment, the housing material may contain polylactic acid (PLA) and polybutylene adipate terephthalate (PBAT), preferably in a weight ratio of PLA:PBAT between 95:5 and 70:30, or about 90:10. Unless otherwise defined, ratios as used herein mean weight ratios (or "weight ratios", "w / w"), where the ratio is calculated with reference to the total weight of the housing components used.

[0101] The materials for the housing may also include PLA, PBAT, PBSA, and / or PBS in the different proportions shown in the table below:

[0102] Table 1. Housing materials. [Table 1]

[0103] Optionally, the housing is 5-100% or 10-100% PLA w / w. Optionally, the housing is 20-90% or 30-80% PBS w / w. Optionally, the housing is 20-100% or 30-90% PBAT w / w. Optionally, the housing is 20-100% or 30-90% PBSA w / w. Optionally, the housing is 5-100% or 10-100% PLA w / w, and one of the following: (i) 20-90% or 30-80% PBS w / w, (ii) 20-100% or 30-90% PBAT w / w, or (iii) 20-100% or 30-90% PBSA w / w.

[0104] As outlined above, the housing of the bolus of this disclosure may be designed in such a way that the active ingredient, i.e., the methane inhibitor, can pass through the housing. This can provide sustainable, controlled release. In one embodiment, the methane inhibitor can flow through the housing material of the bolus of this disclosure. Optionally, the methane inhibitor can diffuse through the housing material.

[0105] Alternatively, the housing may be made of one or more non-adsorbent materials, i.e., materials through which the methane inhibitor does not migrate. Using non-absorbent materials for the housing can help control the release rate of the methane inhibitor(s) in a bolus having one or more openings, in a bolus having a housing capable of forming one or more openings, or in an open-end bolus. For example, in these embodiments, the concentration of the methane inhibitor(s) in the core is not reduced by their absorption into the housing material.

[0106] The bolus housings of this disclosure may be given further functional features, for example, by adding further components to the housing material or by altering the dimensions and properties of the housing. In one embodiment, the bolus housing material of this disclosure comprises one or more excipients. In a preferred embodiment, one or more excipients comprise a plasticizer, a curing agent, and / or a coloring agent.

[0107] In one embodiment, the housing further comprises a compound selected from nucleating agents or stabilizers. In one embodiment, the housing does not contain nucleating agents and / or stabilizers. The thickness of the housing may be selected to contribute to the release rate of the methane inhibitor. That is, a relatively thick housing will have a relatively lower release rate than a relatively thin housing. This is especially true when the housing material is permeable to the methane inhibitor. In one embodiment, the housing may have a material thickness less than about 2 mm, preferably in the range of about 0.3 to 1.8 mm, more preferably in the range of about 0.3 to 1.5 mm. In some embodiments, the housing has a material thickness of less than about 1.5 mm, less than about 1.3 mm, or less than about 1 mm. In some embodiments, the housing has a material thickness greater than about 0.8 mm, greater than about 1 mm, or greater than about 1.1 mm. In some embodiments, the housing has a material thickness of about 0.9 mm. In some embodiments, the housing has a material thickness of about 1.2 mm. In some embodiments, the housing has a material thickness of about 1.5 mm. For boluses including housings with one or more openings, thicker housings, such as housing wall thicknesses of up to approximately 5 mm, may also be applicable.

[0108] Optionally, the core and housing have a weight ratio of approximately 3 to 6:1, approximately 4 to 5:1, or approximately 4.6:1.

[0109] In one embodiment, the housing is configured to be disassembled over a predetermined period of time. This predetermined period can be adjusted, for example, through the material thickness of the housing, the selection of the housing material, or the manufacturing process of the housing.

[0110] In one embodiment, the housing encompasses a cavity in which at least a portion of a core is disposed, wherein the core contains a methane inhibitor such as a methane inhibitor. In another embodiment, the housing does not include an opening and completely encloses the core. In yet another embodiment, the housing completely covers and encloses the core. In one embodiment, the housing encompasses one or more openings as described above.

[0111] The bolus housing of this disclosure may assist in the controlled release of methane inhibitors. For example, the housing may withstand conditions in the lumen over a predetermined period of time. During this time, the housing may protect the core from fluid in the lumen and still facilitate or contribute to the controlled release of methane inhibitors. However, the housing design may allow the housing to disintegrate or decompose over a predetermined period of time. This may help mitigate the adverse effects of device administration to animals and may also ensure that animals can be treated with multiple boluses. For example, a second bolus may be administered at the end of a predetermined period, towards the end of a predetermined period, or after the end of a predetermined period.

[0112] In the aspects described, when the housing is exposed to a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C, the housing becomes permeable to the methane inhibitor, and / or at least a portion of the housing is configured to form one or more openings, allowing the methane inhibitor to exit the bolus through the opening(s). Alternatively, the permeability of the housing to the methane inhibitor may increase when the housing is exposed to a critical minimum temperature, or the escape through the formed openings may increase. In the latter case, the permeability and / or escape of the methane inhibitor increases compared to the permeability / escape from a bolus that is not exposed to at least the aforementioned temperature, e.g., a bolus exposed to a temperature below 28°C.

[0113] A bolus that provides a temperature-responsive release of a methane inhibitor allows the release of the methane inhibitor or an increase in its release rate to a substantial amount only upon exposure of the bolus to the minimum required temperature, thus having an effect on the environment of the bolus. This means that the bolus will release an effective amount of methane inhibitor upon administration to an animal, but not before. Specifically, to ensure that no methane inhibitor is released from the bolus, or that no substantial amount is released, the bolus may be kept at room temperature or in a low-temperature environment prior to administration. Preventing premature release of methane inhibitor from the bolus in this manner may provide several benefits, including: preventing or reducing any loss of methane inhibitor before actual administration; preventing or reducing contamination by potentially aggressive methane inhibitors in the environment / surroundings where the bolus is kept; and protecting farmers, staff, or other people handling the bolus from potential harmful exposure to methane inhibitors.

[0114] In one embodiment, the housing includes one or more openings which are filled with and / or covered with a material which melts, dissolves, or collapses in the rumen of a living animal. Preferably, the housing is made of a material which does not melt, dissolve, or collapse in the rumen of a living animal, preferably over a 24-hour period.

[0115] To enable temperature-dependent release, at least a portion of the bolus, in which an opening may be formed or in which permeability to a methane inhibitor may be increased, may contain or consist of a certain suitable compound or mixture of compounds having properties that may change upon reaching a certain temperature transition point. In one embodiment, a portion of the housing, preferably a portion of the housing in which an opening(s) is formed, is made of a material that melts, dissolves, or disintegrates in the rumen of a living animal.

[0116] In one embodiment, the material to be melted is a compound selected from the group consisting of hydrogels, oleogels, organogels, phase change materials (PCMs), fatty acids, alkanes, alkenes, gelling agents, waxes, L-alanine amino acids, L-alanine amino acid derivatives, poly(methyl methacrylate) (PMMA), (1,3:2,4) dibenzylidene sorbitol (DBS), hydroxystearic acid, paraffin wax, gelatin, 1-tetradecanol, polyethylene glycol, octadecane, nonadecane, eicosane, Pluronic® polymer or mixtures of Pluronic®, emulsifiers, sucrose acetate isobutyrate (SAIB), derivatives of the above, and one or more combinations of the above compounds, wherein the compound or combination of compounds has a melting temperature between 28°C and 42°C, more preferably between 28°C and 35°C.

[0117] The aforementioned melting temperature is the temperature at which a substance changes substantially from a solid to a liquid state at atmospheric pressure.

[0118] In another embodiment, the material to be dissolved is preferably a water-soluble material selected from the group consisting of polymers, polyols, sugars, polyamides, salts, cellulose acetate, polyethylene glycol, methylcellulose, CMC, polyvinyl alcohol, alginates, polyacrylic acid or its salts, polyacrylamide, cellulose ethers, carrageenan, guar, and pectin. "Water-soluble" means that the substance can dissolve in distilled water at a temperature of 20°C.

[0119] In another embodiment, the material to be disintegrated is a compound selected from the group consisting of cellulose, polyhydroxyalkanoate (PHA), poly(butylene succinate-co-adipate) (PBSA), and mixtures of two or more of the above. The lumen environment contains enzymes that can disintegrate or assist in disintegrating the above compounds. When the material disintegrates, the release of the active agent from inside the bolus to outside the bolus will increase. This can be tested as outlined above in the context of quantifying the release of the active agent from inside the bolus.

[0120] In another aspect, the present disclosure relates to a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core comprising a methane inhibitor; and a housing covering at least a portion of the core, wherein at least a portion of the housing is configured to form one or more openings, allowing the methane inhibitor to exit the bolus through the opening(s), wherein the portion of the housing formed by the opening(s) is a temperature-responsive hydrogel, oleogel, organogel, phase change material (PCM), fatty acid, alkane, The material comprises or consists of compounds selected from the group consisting of alkenes, gelling agents and waxes, L-alanine amino acids or L-alanine amino acid derivatives, poly(methyl methacrylate) (PMMA), (1,3:2,4) dibenzylidene sorbitol (DBS), hydroxystearic acid, paraffin wax, gelatin, 1-tetradecanol, polyethylene glycol, octadecane, nonadecane, eicosane, Pluronic® polymer or mixture of Pluronic®, emulsifiers, sucrose acetate isobutyrate (SAIB), the derivatives described above, and one or more combinations of the compounds described above. In some embodiments, the housing surrounds the core.

[0121] In another aspect, the present disclosure relates to a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core containing a methane inhibitor; and a housing covering at least a portion of the core, wherein at least a portion of the housing is configured to form one or more openings, allowing for increased release of the methane inhibitor from within the bolus through the openings(s)(s), wherein the portion of the housing formed by the openings(s)(s) is a temperature-responsive hydrogel, oleogel, organogel, phase change material (PCM), fatty acid, alkali The material comprises or consists of compounds selected from the group consisting of ammonium compounds, alkenes, gelling agents and waxes, L-alanine amino acids or L-alanine amino acid derivatives, poly(methyl methacrylate) (PMMA), (1,3:2,4) dibenzylidene sorbitol (DBS), hydroxystearic acid, paraffin wax, gelatin, 1-tetradecanol, polyethylene glycol, octadecane, nonadecane, eicosane, Pluronic® polymer or mixtures of Pluronic®, emulsifiers, sucrose acetate isobutyrate (SAIB), the derivatives described above, and one or more combinations of the compounds described above. In some embodiments, the housing surrounds the core.

[0122] In another embodiment, the portion of the housing formed by the opening(s) comprises or consists of a compound selected from the group consisting of temperature-responsive hydrogels, oleogels, organogels, phase change materials (PCMs), fatty acids, alkanes, alkenes, gelling agents, and waxes, preferably the compound having a melting temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C.

[0123] In another embodiment, the portion of the housing formed by the opening(s) comprises or consists of a compound selected from the group consisting of temperature-responsive hydrogels, oleogels, organogels, phase change materials (PCMs), fatty acids, alkanes, alkenes, gelling agents, and waxes, preferably the compound being solid at 30°C and liquid at 42°C.

[0124] In a preferred embodiment, the compound is selected from the group consisting of L-alanine amino acid derivatives, poly(methyl methacrylate) (PMMA), (1,3:2,4) dibenzylidene sorbitol (DBS), hydroxystearic acid, paraffin wax, gelatin, 1-tetradecanol, polyethylene glycol (PEG), octadecane, nonadecane, eicosane, Pluronic® polymer or mixture of Pluronic®, emulsifiers, sucrose acetate isobutyrate (SAIB), the derivatives described above, and one or more combinations of the compounds described above.

[0125] The material of the portion of the housing not formed by the opening(s) includes compounds selected from the group consisting of PLA, PCL, PBS, PBAT, PHB, PBSA, wood flour, and one or more combinations of these compounds. Such materials have the property of having a melting temperature higher than the temperature at which the animal's lumen is located. This means that the portion formed by the opening(s) will be allowed to melt at the lumen, while the rest of the housing will remain intact and solid when exposed to the same lumen temperature. Organogel

[0126] In a further aspect, a bolus is provided for administration to ruminants, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: A core comprising a methane inhibitor dispersed in or forming part of one or more hydrogels, oleogels, or organogels; and The housing covers at least a portion of the core.

[0127] Hydrogels, oleogels, and organogels offer a favorable (controlled and / or extended) release profile, are relatively easy to handle, and simplify manufacturing. Furthermore, in at least a preferred embodiment, hydrogels, oleogels, and organogels provide a relatively high level of release at lower loading amounts of the active ingredient compared to equivalent boluses with other carriers (particularly polymer-based carriers), potentially reducing the amount of active agent required per bolus.

[0128] The hydrogels used herein may be temperature-responsive hydrogels. A temperature-responsive hydrogel is understood as a gel containing a crosslinked polymer network in which the swelling agent is water or an aqueous solution. Hydrogels may be based on crosslinked polymers, including, but not limited to, natural polymers, N-isopropylacrylamide polymers, poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) polymers, and poly(ethylene glycol)-biodegradable polyester copolymers.

[0129] Organogels are understood as preparations comprising organic liquids and gelling agent components, which form gels filled with organic liquids including but not limited to polar organic solvents and their aqueous mixtures, ionic liquids, fats, and oils. The thermal stability of the gel can be adapted by selecting the solvent according to its boiling point and, generally, by selecting the components according to their critical phase transition temperature.

[0130] Oleogels, classified as a type of organogel, are understood as preparations whose main components may consist of paraffin oil, fat, or natural oil, usually with the addition of polyethylene, to form so-called isogels, or oils gelled with various additives to form so-called heterogels. Gelling agents used in heterogels include zinc stearate, aluminum stearate, highly dispersed silicon dioxide, and ethylcellulose.

[0131] In some embodiments, the core is a hydrogel. In some embodiments, the core is an oleogel. In some embodiments, the core is an organogel.

[0132] In some embodiments where the core is a hydrogel, oleogel, or organogel, the core comprises a further active agent. In some embodiments where the core is a hydrogel, oleogel, or organogel, the core comprises a further methane inhibitor (preferably lecithin). In some embodiments where the core is a hydrogel, oleogel, or organogel, a combination of a methane inhibitor and a further active agent (preferably lecithin), optionally further containing water, forms a hydrogel, oleogel, or organogel. In some embodiments where the core is a hydrogel, oleogel, or organogel, the core consists of a methane inhibitor, a further methane inhibitor (preferably lecithin), and optionally water.

[0133] In some embodiments where the core is a hydrogel, oleogel, or organogel, the core comprises a phospholipid (preferably lecithin) and / or an acrylate polymer (preferably poly(methyl methacrylate) [PMMA]). In some embodiments where the core is a hydrogel, oleogel, or organogel, the housing surrounds the core. Optionally, Pluronic®, also known as poloxamers, is a class of synthetic block copolymers consisting of hydrophilic poly(ethylene oxide) (PEO) and hydrophobic poly(propylene oxide) (PPO) arranged in an ABA triblock structure, thus giving PEO-PPO-PEO. When mixed with water, concentrated solutions of poloxamers can also form hydrogels, which may also be the preferred compounds described. Increasing the temperature to certain critical levels can affect the hydrogen bonding between polyoxyethylene and water molecules, altering the cohesive forces of the Pluronic® / poloxamer components and potentially increasing permeability.

[0134] Sucrose acetate isobutyrate (SAIB), prepared by esterification of sucrose with acetic acid and isobutyric anhydride, is known to be an acceptable emulsifier in food preparation and therefore may exhibit particularly tolerable properties when used in animals.

[0135] In some embodiments, the portion of the housing formed by the opening(s) contains or comprises a compound whose properties change to allow increased permeability at a critical temperature, which is the melting point of the compound. This is, for example, when oil and / or wax is included in the portion of the housing formed by the opening(s) and / or the housing to be permeable to a methane inhibitor.

[0136] Those skilled in the art know that when the portion of the housing formed by the opening(s) contains or consists of polyethylene glycol, the critical temperature for allowing increased permeability depends on the PEG or PEG mixture used. Those skilled in the art can select a suitable PEG component to achieve the desired critical emission temperature. PEG can also be suitably used in the form of a PEG gel, i.e., a gel containing PEG as a functional phase.

[0137] Phase change materials (PCMs) are materials or compounds that can melt or solidify at certain temperatures. Thermal energy is absorbed or released by changing the state of the material. Phase change materials may be solid at room temperature and may soften when they reach a specific temperature. This can include the material becoming more permeable. Bio-based PCMs include glycols, alcohols, esters, and fatty acids. Specifically, fatty acids, especially saturated fatty acids, such as palmitic acid and stearic acid, are preferred PCMs, and these may be derived from vegetable oils and animal fats, such as palm oil or coconut oil. Further biomedically, and therefore veterinarily, applicable phase change materials are known in the art and include, but are not limited to, 1-tridecanol, 1-tetradecanol, 1-pentadecanol, decanoic acid, or lauric acid.

[0138] The largest opening formed on the housing when the housing is exposed to the rumen of a living animal and / or at least to the temperature of the lumen or physiological temperature may preferably have a diameter of up to 1 mm to ensure that the release of the methane inhibitor through the opening remains controlled and sustained.

[0139] In one embodiment, the largest opening formed on the housing when the housing is exposed to the lumen of a living animal has a maximum diameter of 2 mm. In one embodiment, the largest opening formed on the housing when the housing is exposed to the lumen of a living animal has a maximum diameter of 1 mm.

[0140] In one embodiment, when the housing is exposed to a temperature preferably 42°C relative to the temperature present in the rumen of a living animal, the largest opening formed on the housing has a maximum diameter of 2 mm. In one embodiment, when the housing is exposed to a temperature preferably 42°C relative to the temperature present in the rumen of a living animal, the largest opening formed on the housing has a maximum diameter of 1 mm.

[0141] In another embodiment, the largest opening formed on the housing when the housing is exposed to a temperature of at least 38°C has a maximum diameter of 2 mm. In another embodiment, the largest opening formed on the housing when the housing is exposed to a temperature of at least 38°C has a maximum diameter of 1 mm. The size of the formed pores may be deterministic to the release rate of the methane inhibitor from the bolus, where the methane inhibitor is released through the formed pores.

[0142] Discharge mechanisms provided by boluses that involve temperature-enhanced or temperature-dependent release of the methane inhibitor contained within the bolus are useful when delayed release is required. In this context, "delayed" should be understood as delaying or substantially slowing down the release of the methane inhibitor from the bolus until after administration to the ruminant, i.e., after placing the bolus in the animal's rumen. This can be advantageous in handling the bolus. Here, the bolus releases a relatively small amount or even none of the methane inhibitor contained within it when handled outside the rumen environment, and only begins releasing the methane inhibitor after administration to the animal's rumen, or begins releasing it at a higher, for example, effective rate. The benefits provided by this bolus modification include protecting farmers or other staff handling the bolus from methane inhibitors such as bromoform, which may have health hazards or irritating effects on humans when in direct contact with humans.

[0143] The release rate of methane inhibitor from the bolus through one or more openings may increase when the housing is exposed to temperatures at least physiological or lumen temperatures. This allows for increased escape of methane inhibitor from the bolus through the opening(s).

[0144] In one embodiment, the bolus of the present disclosure comprises the methane inhibitor bromoform and is adapted to reach a maximum release rate of approximately 0.1 to approximately 0.5 g per day, more preferably approximately 0.2 g per day. Such a release rate can provide a sustained release of a haloform such as bromoform. Bolus having such a release rate is suitable for use, for example, in livestock such as cattle and sheep.

[0145] To achieve a desirable release rate, the concentration of the methane inhibitor, such as haloform, or the thickness of the housing material may also be adjusted in smaller livestock. Furthermore, to achieve a desirable release rate, the size of the opening formed in the bolus of this disclosure by administration to the rumen may also be adjusted in smaller livestock. Furthermore, to achieve a desired release rate of the methane inhibitor, the overall polarity of the carrier material that can be mixed with the methane inhibitor may be adjusted to achieve a desired affinity for the methane inhibitor mixed with it. However, in an alternative embodiment, the methane inhibitor may be provided in a substantially pure form, for example, not mixed with a carrier.

[0146] In one embodiment of any bolus including a housing surrounding a methane inhibitor, the bolus is adapted to release the methane inhibitor over a period of at least two months. In a preferred embodiment, the bolus is adapted to release the substance over a period of at least six months, for example, at least seven, eight, nine, or at least ten months.

[0147] In one embodiment of any bolus described herein, the methane inhibitor is selected from haloforms.

[0148] In one embodiment, the release rate of the methane inhibitor from the bolus through one or more openings in the housing increases when the housing is exposed to temperatures between 28°C and 42°C, preferably between 28°C and 42°C, compared to the release rate of the bolus when exposed to a temperature of 20°C. In other words, there is an increased escape of the methane inhibitor from the bolus through the opening(s) when the bolus is inside the lumens of a living animal. In one embodiment, the release rate of the methane inhibitor from the bolus through one or more openings increases when the housing is exposed to temperatures of at least 38°C, allowing for an increased escape of the methane inhibitor from the bolus through the opening(s).

[0149] The aforementioned increase in the release rate of the methane inhibitor from the bolus through one or more openings is influenced by the diameter of the opening or pore formed. This is, for example, no more than 1 mm to maintain controlled and sustained release, as described above.

[0150] A preferred embodiment of the first aspect of this disclosure relates to a bolus of the first aspect that can be produced by performing the following steps: (i) Provision of a housing, wherein the housing material preferably comprises PLA and PBAT in a wt% ratio of about 90:10 PLA:PBAT, and preferably has a wall thickness of about 0.5 to 2 mm; (ii) Fill the housing with at least 20g of steel balls; (iii) Mixing ethylcellulose with tribromomethane; (iv)(iii) is mixed with hydroxypropyl methylcellulose, thereby the resulting mixture preferably having a weight ratio of about 3:1:1, comprising tribromomethane, ethylcellulose, and hydroxypropyl methylcellulose; (v) sealing the housing by spin-welding a cap onto it; Here, the housing is configured to increase its permeability to the methane inhibitor when the housing is exposed to the lumen of a living animal; and / or, here, at least a portion of the housing is configured to form one or more openings when the housing is exposed to the lumen of a living animal, allowing the methane inhibitor to exit the bolus through the opening(s).

[0151] In a further aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core comprising a methane inhibitor in microencapsulated particles, the microencapsulated particles dispersed within a carrier; and optionally a dispersant, as well as a housing covering at least a portion of the core.

[0152] In a further embodiment, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core comprising a methane inhibitor in microencapsulated particles, the microencapsulated particles dispersed within a carrier; and a dispersant, as well as a housing covering at least a portion of the core.

[0153] Encapsulating methane inhibitors as microencapsulated particles can offer several advantages, which will become clearer below. For example, since microcapsules can be uniformly dispersed within a bolus, microencapsulation facilitates the uniform and sustained release of methane inhibitors from the microcapsules contained within the bolus. Furthermore, encapsulating methane inhibitors prevents or reduces their direct contact with other bolus components, protecting these components from potentially aggressive activity as methane inhibitors.

[0154] As used herein, “dispersant” is a substance added to a mixture or suspension of solid or even liquid particles in a carrier in order to improve the separation of particles and to prevent their sedimentation or agglomeration. Suitable dispersants include ionic (e.g., anionic) and nonionic surfactants, polyethylene glycol and its derivatives, glucosides, and others, and those skilled in the art will know suitable compounds and polymers to be used as dispersants.

[0155] In one embodiment, microencapsulated particles are produced by microencapsulation in at least one encapsulating agent, preferably, wherein the encapsulating agent is selected from the group consisting of polymers, surfactants, emulsifiers, gelatin-sorbitol mixtures and gelatin-starch syrup, and mixtures thereof, more preferably, wherein the encapsulating agent is selected from the group consisting of PLA, PBSA, PBS, PHBV, PVA, PBAT, PCL, PDLA, gelatin-sorbitol mixtures and gelatin-starch syrup. In particular, since certain degrees of brittleness of the encapsulating compound are not detrimental to stability at small microcapsule sizes, a wide variety of, for example, polymer compounds can be used for microencapsulation due to the small size of the microcapsules.

[0156] Microcapsulation methods are well known in the art and will be obvious to those skilled in the art. For example, the method described by Aida et al. (1989) can be applied to microcapsulate methane inhibitors such as haloform (Aida et al., “Practical Application of Microcapsulation for Toxicity Studies Using Bromodichloromethane as a Model Compound”, JOURNAL OF THE AMERICAN COLLEGE OF TOXICOLOGY, Volume 8, Number 6, 1989). In one embodiment, microcapsulated particles are produced by microcapsulation in gelatin-starch syrup.

[0157] When an oil phase is used and the substance to be encapsulated is mixed therein, the oil phase may preferably contain further stabilizers, such as emulsifiers, to assist in particle formation. Therefore, in one embodiment, the oil phase used in the process of forming microencapsulated particles may contain at least one stabilizer, preferably at least one emulsifier, and more preferably the oil phase contains lecithin.

[0158] In one embodiment, microencapsulated particles may be microencapsulated using interfacial polymerization. During interfacial polymerization, polymerization occurs at the interface between two immiscible phases, such as two liquids, resulting in a polymer residing in the interfacial layer. Preferred process modifications of interfacial polymerization for encapsulation are known in the art, as described, for example, by Song et al. (2017) (Song et al., "Recent progress in interfacial polymerization." Materials Chemistry Frontiers 1.6 (2017):1028-1040), and will be obvious to those skilled in the art. In one embodiment, microencapsulated particles may be microencapsulated without using interfacial polymerization.

[0159] Porous carrier materials may be suitable for dispersing microcapsules. For example, bromoform, as an exemplary methane inhibitor, may not be directly compatible with hydrophilic carrier materials, but microcapsules containing bromoform can be dispersed within a hydrophilic carrier. However, it should be assumed that once bromoform is released from the microcapsule, for example, through the porous structure of the carrier, it can still escape from the bolus.

[0160] In a further aspect, the present disclosure provides a bolus for administration to ruminants, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a carrier and microencapsulated particles, wherein the microencapsulated particles contain a methane inhibitor, and the microencapsulated particles are dispersed within the carrier, preferably wherein the carrier has a porous structure. Such a porous structure may contain small or fine spaces or pores through which liquids such as air and rumen fluid can pass. Thus, microencapsulated particles dispersed within a carrier having a porous structure can be contacted by rumen fluid passing through the pores, facilitating the delivery of the methane inhibitor from the microencapsulated particles to the rumen fluid.

[0161] In one embodiment, microencapsulated particles are microencapsulated using a hydrophilic material, gelatin, zein, methylcellulose and poly(N-isopropylacrylamide) (PNIPAM) microgel, starch, cyclodextrin, and compounds selected from the group consisting of two or more combinations of the aforementioned compounds.

[0162] For example, cyclodextrins can be used, particularly due to their beneficial ability to retain hydrophobic compounds within an encapsulated core. For instance, poly(N-isopropylacrylamide) (PNIPAM) microgels are widely used in biomedical applications and contain colloidal particles that form the microgel.

[0163] Microencapsulating methane inhibitors allows the use of various different methane inhibitors with various different bolus carrier components in which the microcapsules are dispersed. This is because, since the methane inhibitor is not loaded directly into the carrier but as the contents of the microcapsule, the compatibility of the carrier with the methane inhibitor is not particularly required in order to load as much methane inhibitor as possible. In one embodiment, the carrier comprises a compound selected from the group consisting of silica, cellulose and activated carbon, gelatin, chitosan, poly(lactic acid-coglycolic acid) (PLGA), cyclodextrin, collagen, polyalpha-hydroxyesters, hydroxyalkanoates and dioxanes, starch, gluten, zein, polyethylene, polypropylene, polyamide, polyethylene terephthalate, and ethylene vinyl acetate.

[0164] Similar to the exemplary housing materials described herein, the microencapsulated and carrier compounds used in the boluses of this disclosure may also be biodegradable. However, this is not a requirement. Rather, it is preferable that the components used in the boluses described herein should not be harmful to animal health or the environment, especially when accumulated in larger quantities. Therefore, non-biodegradable bolus components are also acceptable for the boluses of this disclosure.

[0165] Furthermore, the microparticle production mechanism ensures a substantially homogeneous structure for all particles. For example, they are approximately the same size and surrounded by an encapsulating agent of approximately the same thickness. Therefore, the release rate of sustained, long-term release from all microcapsules will be approximately the same. In one embodiment, the microencapsulated particles have an average diameter of 50 nm to 2 mm, preferably 1 to 1000 μm. The diameter of the microencapsulated particles can be determined, for example, by sieve analysis, using different standard sieve sizes. Moreover, scanning electron microscopy of the particles can be applied, which allows for further determination of the particle shape and surface morphology.

[0166] In one embodiment, the bolus is configured to release a methane inhibitor over a period of at least six months. In one embodiment, the bolus is configured to release a methane inhibitor over a period of up to six months. In one embodiment, the bolus is configured to release a methane inhibitor over a period of at least four months. In one embodiment, the bolus is configured to release a methane inhibitor over a period of at least two months. No housing

[0167] In a further aspect, the present disclosure provides a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises a core, the core comprises a methane inhibitor and a carrier, and the bolus does not comprise a housing.

[0168] In some embodiments, preferably in embodiments where the bolus does not include a housing, the carrier is hydrophobic. In some embodiments, preferably in embodiments where the bolus does not include a housing, the carrier consists of one or more hydrophobic materials. In some embodiments, preferably in embodiments where the bolus does not include a housing, the carrier has a melting point of at least 60°C, preferably at least 75°C.

[0169] In some embodiments, preferably in embodiments where the bolus does not include a housing, the bolus does not include carboxylate glass. In some embodiments, preferably in embodiments where the bolus does not include a housing, the bolus does not include glass.

[0170] In some embodiments, preferably in embodiments in which the bolus does not include a housing, the bolus does not contain a seaweed extract concentrated with bromoform. In some embodiments, preferably in embodiments in which the bolus does not include a housing, the bolus does not contain a seaweed extract.

[0171] In some embodiments where the bolus does not include a housing, the bolus does not contain hydrophobic fumed silica. In some embodiments where the bolus does not include a housing, the bolus does not contain silica.

[0172] In some embodiments where the bolus does not include a housing, the carrier is made of polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), polylactic acid, poly-d-lactic acid, poly-L-lactic acid, poly-D,L-lactic acid (PDLLA), polylactide-co-glycolide, lignin, polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), epoxy chain extender, magnesium silicate, cellulose material, ethylcellulose, hydroxypropyl methylcellulose (HPMC), fumed silica, gelatin, wax, castor wax, paraffin wax, silica, microcrystalline wax, methylcellulose, starch, polyethylene glycol, polyvinyl alcohol, hydroxyethylcellulose, carboxymethylcellulose, Solplus, poly(acrylic acid), poly(vinylpyrrolidone), poly(vinyl alcohol), poly(acrylamide), poly(2-hydroxypropyl methacrylamide), poly(N,N-dimethyl acrylate) The material comprises one or more materials selected from the list consisting of poly([2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide), poly(2-(methacryloyloxy)ethyl phosphorylcholine), poly(carboxybetaine methacrylamide), poly(ethylene glycol), poly(ethyleneimine), poly(sarcosine), poly(2-methyl-2-oxazoline), polyaminoesters, polyesteramides, polyphosphoesters, poly(l-lysine), poly(l-proline), polyphosphazenes, dextran, sodium alginate, gelatin, agarose, carrageenan, guerlain, xanthan gum, urea, sucrose, their derivatives, combinations thereof, and their copolymers.In some embodiments where the bolus does not include a housing, the carrier is made of polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), polylactic acid, poly-d-lactic acid, poly-L-lactic acid, poly-D,L-lactic acid (PDLLA), polylactide-co-glycolide, lignin, polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), epoxy chain extender, magnesium silicate, cellulose material, ethylcellulose, hydroxypropyl methylcellulose (HPMC), fumed silica, gelatin, wax, castor wax, paraffin wax, silica, microcrystalline wax, methylcellulose, starch, polyethylene glycol, polyvinyl alcohol, hydroxyethylcellulose, carboxymethylcellulose, Solplus, poly(acrylic acid), poly(vinylpyrrolidone), poly(vinyl alcohol), poly(acrylamide), poly(2-hydroxypropyl methacrylamide), poly(N,N-dimethylacrylic It consists of one or more materials selected from the list consisting of amides, poly([2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide), poly(2-(methacryloyloxy)ethyl phosphorylcholine), poly(carboxybetaine methacrylamide), poly(ethylene glycol), poly(ethyleneimine), poly(sarcosine), poly(2-methyl-2-oxazoline), polyaminoesters, polyesteramides, polyphosphoesters, poly(l-lysine), poly(l-proline), polyphosphazenes, dextran, sodium alginate, gelatin, agarose, carrageenan, guerlain, xanthan gum, urea, sucrose, their derivatives, combinations thereof, and copolymers thereof.In some embodiments where the bolus does not include a housing, the carrier consists of PLA, PBSA, PBS, PHBV, PVA, PBAT, PCL, PDLA, epoxy chain extenders, magnesium silicate, cellulose materials, ethylcellulose, hydroxypropyl methylcellulose (HPMC), fumed silica, gelatin, wax, caster wax, paraffin wax, silica, and combinations thereof. In some embodiments where the bolus does not include a housing, the carrier includes microcrystalline wax. In some embodiments where the bolus does not include a housing, the carrier consists of microcrystalline wax.

[0173] As used herein, “carrier” is a compound that can be mixed with methane inhibitors and / or other active agents without altering the chemical structure of the methane inhibitor and / or other active agents. Preferably, when used in the bolus of this disclosure, the carrier delays the release of the methane inhibitor and / or other active agent from the bolus.

[0174] For example, the support may contain at least one polar functional group.

[0175] The functional groups covalently linked to the support can be selected from the group consisting of esters, fatty acids, fatty alcohols, carbonyls, and fatty amines. Without being constrained by theory, such modified support can interact with the methane inhibitor via polar functional groups, and partly possibly via hydrogen bonding.

[0176] A wide variety of materials may be suitable for use as a support in the bolus of this disclosure, and the following examples are not limited. For example, the support may be selected from the list of waxes, myristic acid, stearic acid, steryl alcohols, cetyl alcohols, cetosteryl alcohols, or combinations thereof. The support may be a waxy substance. For example, the support may be selected from the list of beeswax, paraffin wax, PEG4000, carnauba, castor wax, candelilla, jojoba, or lanolin, or combinations thereof. The support may comprise a mixture of two or more components, for example, a relatively nonpolar substance and at least one relatively polar substance. As a result, the overall polarity of the support may be adjusted to achieve a desired affinity for the methane inhibitor. This may be used to achieve a desired release rate for the methane inhibitor. 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 have relatively high amounts of polar functional groups).

[0177] In one embodiment, the bolus may be adapted to exhibit an release rate into the rumen between 0.02 g and 2 g per day, preferably approximately 0.1 to 0.5 g of bromoform per day. When the bolus exhibits such a release rate for a methane inhibitor (e.g., a haloform such as bromoform), this can reduce methane production. The release rate of the methane inhibitor into the rumen may increase over time; that is, the release rate starts from zero upon administration to the animal and increases to a maximum due to several factors. However, the foregoing should not be seen as limiting, and other release rates are assumed to be within the scope of this disclosure.

[0178] The bolus supports described herein may have a melting point below the boiling point of the methane inhibitor. This may be useful because the support can melt and mix with the methane inhibitor without substantial loss of the methane inhibitor due to evaporation. Furthermore, having a melting point above 37°C, more preferably above 40°C, can assist the support in stabilizing the methane inhibitor when the bolus is in the lumen. This means that the bolus core will not melt in the lumen, which in some cases may have temperatures of up to approximately 40°C. This may be useful, for example, for boluses including a housing, in order to control the release of the methane inhibitor, including its movement through the material forming the housing.

[0179] In addition, the support may include powdered activated carbon, zeolite or bentonite, elemental zinc or zinc oxide. Preferably, high-density materials such as metal (preferably steel) pieces may be included in the support. Additional components may be used to achieve the desired density for the core and / or bolus.

[0180] It should be understood by those skilled in the art that other carriers and / or core components may be selected or used depending on the application. It is assumed that certain carriers may be selected to provide a desired release profile for the methane inhibitor or, alternatively, to provide desired physical properties, density, or volume, etc., of the core material. In one embodiment, a carrier contained in a bolus, which also includes a housing, may have a relatively high affinity for the methane inhibitor compared to the affinity of the housing to the methane inhibitor. This can be achieved, for example, by the relative polarity of the materials forming the carrier and the housing and by appropriately matching these materials to the methane inhibitor.

[0181] In one embodiment, the core is composed of polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), polylactic acid, poly-d-lactic acid, poly-L-lactic acid, poly-D,L-lactic acid (PDLLA), polylactide-co-glycolide, lignin, polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), and talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), epoxy chain extender, magnesium silicate, cellulose material, ethylcellulose, hydroxypropyl methylcellulose (HPMC), fumed silica, gelatin, wax, castor wax, paraffin wax, silica, hydrophilic silica, microcrystalline wax, methylcellulose, starch, polyethylene glycol, polyvinyl alcohol, hydroxyethylcellulose, carboxymethylcellulose, solplus, poly(acrylic acid), poly(vinylpyrrolidone), poly(vinyl alcohol), poly(acrylamide), poly(2-hydroxypropyl methacrylamide), poly(N,N-dimethylacrylamide), poly([2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide), poly(2-(methacryloyloxy)ethyl phosphorylcholine), poly(carboxybetaine methacrylamide), poly(ethylene glycol), poly(ethyleneimine), poly(sarcosine), poly(2-methyl-2-oxazoline), polyaminoester, polyesteramide, polyphosphoester, poly(l-lysine), poly(l-proline), polyphosphazene, dextran, sodium alginate, gelatin, agarose, carrageenan, guerlain, xanthan gum, urea, sucrose, beeswax, polyethylene glycoside The core comprises one or more materials selected from the list consisting of polyacrylic acid (PEG), sodium starch glycolate, croscarmellose sodium, crospovidone, carboxymethylcellulose (CMC), hydroxyethylcellulose (HEC), carrageenan, guar gum, xanthan gum, sodium alginate, locust bean gum, polyvinylpyrrolidone (PVP), polyvinylpyrrolidone-vinyl acetate copolymer (PVP / VA), polyacrylic acid and / or copolymer variants thereof, polyisobutylene, vinyl ethyl acetate (EVA), functional waxes having a melting point less than about 120°C, their derivatives, combinations thereof, and their copolymers. In one embodiment, the core comprises one or more materials selected from the list consisting of polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), polylactic acid, poly-d-lactic acid, poly-L-lactic acid, poly-D,L-lactic acid (PDLLA), polylactide-co-glycolide, lignin, polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), epoxy chain extender, magnesium silicate, cellulose material, ethylcellulose, hydroxypropyl methylcellulose (HPMC), fumed silica, gelatin, wax, castor wax, paraffin wax, silica, microcrystalline wax, methylcellulose, starch, polyethylene glycol, polyvinyl alcohol, hydroxyethylcellulose, carboxymethylcellulose, solplus, poly(acrylic acid), poly(vinylpyrrolidone), poly(vinyl alcohol), poly(acrylamide), poly(2-hydroxypropyl methacrylamide), poly(N,N-dimethylacrylamide), poly([2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide), poly(2-(methacryloyloxy)ethyl phosphorylcholine), poly(carboxybetaine methacrylamide), poly(ethylene glycol), poly(ethyleneimine), poly(sarcosine), poly(2-methyl-2-oxazoline), polyaminoesters, polyesteramides, polyphosphoesters, poly(l-lysine), poly(l-proline), polyphosphazenes, dextran, sodium alginate, gelatin, agarose, carrageenan, guerlain, xanthan gum, urea, sucrose, beeswax, polyethylene glyco It consists of one or more materials selected from the list comprising: PEG (PEG), sodium starch glycolate, croscarmellose sodium, crospovidone, carboxymethylcellulose (CMC), hydroxyethylcellulose (HEC), carrageenan, guar gum, xanthan gum, sodium alginate, locust bean gum, polyvinylpyrrolidone (PVP), polyvinylpyrrolidone-vinyl acetate copolymer (PVP / VA), polyacrylic acid and / or copolymer variants thereof, polyisobutylene, vinyl ethyl acetate (EVA), functional waxes having a melting point less than about 120°C, their derivatives, combinations thereof, and their copolymers.

[0182] In one embodiment, the core comprises a compound selected from the group consisting of polylactic acid (PLA), poly(butylene succinate-co-butylene adipate (PBSA), polybutylene succinate (PBS), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), polybutylene adipate terephthalate (PBAT), polycaprolactone (PCL), poly(D,L-lactic acid) (PDLA), epoxy chain extenders, magnesium silicate, cellulose materials, ethylcellulose, hydroxypropyl methylcellulose (HPMC), fumed silica, gelatin, wax, caster wax, paraffin wax, silica, and combinations thereof.

[0183] Those skilled in the art will know of further compounds, specifically further polymers, preferably biodegradable polymers, that can be suitably used as core components of the bolus in this embodiment.

[0184] In one embodiment, the bolus of the disclosed herein comprises hydrophobic fumed silica. Preferably, such fumed silica is amorphous or consists of or comprises hydrophobic fumed silica particles (HFSPs). In one preferred embodiment, the average particle size of the hydrophobic fumed silica is between 5 nm and 15 nm. For example, the bolus may contain at most 10 wt%, at most 8 wt%, or at most 5 wt% of the hydrophobic fumed silica. Preferably, the bolus contains at most 5 wt% of the hydrophobic fumed silica, where the methane inhibitor is bromoform. In one embodiment, hydrophobic fumed silica is silica that can be produced by contacting silica with a hydrophobic silane, preferably a compound selected from the group consisting of dimethyldichlorosilane (DDS), methylacrylsilane, octylsilane, octamethylcyclotetrasiloxane, hexadecylsilane, octylsilane, methylacrylsilane, polydimethylsiloxane, hexamethyldisilazane (HMDS), silicone oil, silicone oil plus aminosilane, HMDS plus aminosilane, organic phosphate, HMDS (hexamethyldisilazane), and combinations of the above compounds.

[0185] In one embodiment, the bolus of the present disclosure described herein comprises hydrophilic silica.

[0186] The core of a housing-less bolus described herein may be selected to provide sufficient durability of the case-less bolus in a lumen environment. In one embodiment, the bolus has a Shore D hardness of at least 20. In one embodiment, the bolus may have a Shore D hardness of at least 40. The Shore D hardness may be adjusted, for example, through the selection of the core material or the core manufacturing process. It will be obvious to the average person skilled in the art that the bolus hardness may be selected so that the bolus can persist in a lumen environment and withstand physical and chemical influences. In such embodiments, a bolus (without housing) having a Shore D hardness of less than 20 may result in a bolus that is too soft. This may prevent the administration of the bolus to animals, or it may otherwise be damaged or prematurely decomposed before the entire amount of methane inhibitor can be administered.

[0187] Methods for determining Shore D hardness are well known in the art and will be obvious to those skilled in the art. For example, this can be done by using a durometer, which determines Shore D hardness by piercing a sample with the feet of a durometer indenter under a specified spring force.

[0188] The described bolus may function without a housing, but the core of a bolus without a housing may be partially or completely coated. Therefore, in a further embodiment, the bolus may include a core containing a methane inhibitor (preferably a haloform such as bromoform); and a coating covering at least some portion of the core, or preferably the entire core; where the bolus is configured to release the methane inhibitor. A coating layer thickness of less than 2 mm is preferred to allow the methane inhibitor, such as a haloform, to permeate outward from the core material at an optimal rate.

[0189] In one embodiment, the bolus is in the form of a pellet, pill, lozenge, or tablet. The size and shape of the pellet, pill, lozenge, or tablet may be suitably selected by an average person skilled in the art to suit the dose to be administered, the intended duration of administration, and the size of the target animal. For example, a larger pellet, pill, lozenge, or tablet size may be selected for larger animals such as cattle, while a smaller pellet, pill, lozenge, or tablet size may be preferred for smaller ruminants such as sheep.

[0190] Bolus in the form of pellets, pills, lozenges, or tablets may be small boluses, such as boluses having a length of about 1 to 5 cm. Animals may also be administered multiple such small boluses simultaneously or subsequently. For example, animals may be administered multiple such boluses mixed into animal feed, i.e., used as feed additives.

[0191] In one embodiment, the bolus is configured to dissolve in the rumen of a ruminant over a period of less than 48 hours. In a preferred embodiment, the bolus is configured to dissolve in the rumen of a ruminant over a period of less than 12 hours, more preferably less than 6 hours, and even more preferably less than 2 hours. Those skilled in the art will know how to select the coating as size and possibility for the bolus of this disclosure in order to obtain the aforementioned dissolution periods.

[0192] Whether and how quickly a bolus dissolves in the rumen of an animal can be tested, for example, by in vitro testing. This can be done by placing the bolus in a solution with conditions that simulate the rumen environment and determining whether and when the bolus dissolves, i.e., partially or completely disintegrates over time. For in vitro testing, the bolus can be placed, for example, in a tank or vessel containing a known volume (e.g., 1 liter) of phosphate buffer (pH: 6.5, 0.02 M) at 40°C. Optionally, the solution containing the bolus may be stirred to simulate stirring in the rumen. Further active agents

[0193] In a further aspect, a bolus is provided for administration to ruminants, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: core; Housing that covers at least a portion of the core: Here, the core comprises at least one methane inhibitor and at least one further active agent.

[0194] In some embodiments, the housing completely surrounds the core.

[0195] In some embodiments, further active agents are selected from the group consisting of methane inhibitors, hydrogen sequestering agents, anti-inflammatory agents, analgesics, anthelmintics, nonsteroidal anti-inflammatory drugs (NSAIDs), antibiotics, growth promoters, milk secretion promoters, sustained release enhancers, antibacterial agents, ketosis inhibitors, mineral / element / vitamin supplements, and combinations thereof. In some embodiments, further active agents are selected from the group consisting of methane inhibitors, hydrogen sequestering agents, nonsteroidal anti-inflammatory drugs (NSAIDs), anthelmintics, and ketosis inhibitors. In some embodiments, further active ingredients are selected from methane inhibitors, antibiotics, and ketosis inhibitors.

[0196] In some embodiments, further methane inhibitors include haloforms (e.g., haloforms other than bromoform if the first methane inhibitor is bromoform), monensin, phospholipids (e.g., lecithin), and fatty acids (e.g., lauric acid, myristic acid, and linoleic acid); plant extracts or derivatives containing tannins, oils, and essential oils; fumaric acid (e.g., fumaric acid and sodium fumarate), acrylic acid (e.g., sodium acrylate), statins (e.g., atorvastatin and simvastatin), sulfur-containing salts (e.g., sulfuric acid and sodium sulfate), nitric acid (e.g., potassium nitrate, calcium nitrate, calcium ammonium nitrate, and sodium nitrate), malic acid, C 6-14 Fatty acids (preferably C 6-12The group is selected from fatty acids (e.g., apronic acid, enantioic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid), unsaturated fatty acids (e.g., alpha-linolenic acid, stearidonic acid, eicosapentaenoic acid, cervic acid, linoleic acid, linole-elaidic acid, gamma-linolenic acid, dihomo-gamma-linolenic acid, arachidonic acid, docosatetraenoic acid, palmitoleic acid, vaccenic acid, pauric acid, oleic acid, elaidic acid, gondic acid, erucic acid, nervonic acid, meadic acid), and lipids (e.g., fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, sterols, prenolic acid, and saccharolipids). In some embodiments, further methane inhibitors are selected from the group consisting of haloforms (e.g., haloforms other than bromoform if the first methane inhibitor is bromoform), monensin, phospholipids (e.g., lecithin), and fatty acids (e.g., lauric acid, myristic acid, and linoleic acid). In some embodiments, the further methane inhibitor is not 3-nitrooxypropanol (3-NOP). In some embodiments, the hydrogen sequestering agent is fumaric acid, sodium fumarate, phenol compounds, phloroglucinol, gallic acid, resorcinol, catechol, hydro The anti-inflammatory agent / analgesic / NSAID is selected from the group consisting of quinones and pyrogallol. In some embodiments, the anti-inflammatory agent / analgesic / NSAID is selected from nonsteroidal anti-inflammatory drugs (e.g., aspirin, ibuprofen, ketoprofen, carprofen, meloxicam, robenacoxib, firocoxib, mavacoxib, and flunixin), corticosteroids, alpha-2 antagonists, ketamine, and opioid receptor agonists (e.g., tramadol). In some embodiments, the anti-inflammatory agent / analgesic / NSAID is selected from meloxicam and ketoprofen.In some embodiments, the anthelmintic is benximidazole (e.g., mebendazole, flubendazole, fenbendazole, oxfendazole, oxybendazole, albendazole, albendazole sulfoxide, thiabendazole, thiophanate, febantel, netobimine, and triclabendazole, netobimine, albendazole, and triclabendazole), imidazothiazole (e.g., rebamizole), tetrahydropyrimidine (e.g., pyrantel tartarate or embonate, and oxantel), macrocyclic lactone (e.g., ivermectin, abamectin, doramectin, eprinomectin, selamectin, The anthelmintic is selected from the group consisting of milbemycin oxime (and moxidectin), salicylanilide (brothianide, crioxanide, closantel, niclosamide, oxyclozanide, and rafoxanide), substituted phenols (e.g., bithionol, disophenol, hexachlorophene, niclophorane, meniclophorane, and nitroxynyl), aromatic amide diamphenetides (e.g., diamphenetides), praziquantel, epsiplantel, aminoacetonitrile derivatives, cyclic octadepsipeptides (e.g., emodepside), spiroindole (e.g., delkantel), piperazine, chlorthrone, bunamidine, and nitroscanates. In some embodiments, the anthelmintic is albendazole. In some embodiments, the antibiotic is selected from the group consisting of meloxicam, ketoprofen, penicillin, tetracycline, macrolide, monensin, ceftiofur, florfenicol, tilmicosin, enrofloxacin, and tulathromycin.In some embodiments, antimicrobial agents include tetracyclines (e.g., chlortetracycline, oxytetracycline, doxycycline, tetracycline), amphenicol (e.g., florfenicol, thiamphenicol), penicillins and clavulanic acid (e.g., amoxicillin, ampicillin, cloxacillin, pentamate, procain benzylpenicillin, phenoxymethifenicillin), cephalosporins (e.g., cephalonium, cephalexin, cephaprin, cefoperazone, cefquinom, and ceftiofur), lincomamides (e.g., lincomycin), and s The group consists of sulfonamides, trimethoprims, macrolides (e.g., gamithromycin, tildipyrosin, tilmicosin, tulathromycin, tyrosin, and tilvalosin), aminoglycosides (e.g., dihydrostreptomycin, apramycin sulfate, flamycin, neomycin, paromomycin, streptomycin, and spectinomycin), fluoroquinolones (e.g., enrofloxacin, marbofloxacin, and danofloxacin), polymyxins (e.g., colistin), pleuromucilines (e.g., thiamrin), and chloramphenicol. In some embodiments, the ketosis inhibitor is monensin. In some embodiments, the mineral / element / vitamin supplement is selected from the group consisting of copper, cobalt, selenium, manganese, magnesium, sodium and chloride, potassium, zinc, iodine, sulfur, chromium, vitamin A, vitamin E, vitamin D3, and combinations thereof. In some embodiments, further active agents are selected from the group consisting of monensin, phloroglucinol, albendazole, ketoconazole, lecithin, and combinations thereof.

[0197] In particular, the inclusion of multiple methane inhibitors may be beneficial if they act on one or more different pathways, enzymes, and organisms of methane-producing organisms in the rumen. The inclusion of additional active agents to improve the side effects of methane inhibitors may be beneficial. For example, if a methane inhibitor can lead to ketosis, the inclusion of a ketosis inhibitor in the bolus may be particularly useful. The inclusion of additional active ingredients to improve the release profile of methane inhibitors may be beneficial. Generally, a more sustained-release profile is preferred. For example, the inclusion of additional active ingredients that result in a reduced need for administration through the administration of at least two active ingredients at once is desirable.

[0198] In addition to methane, hydrogen gas can also be produced in the lumen. Although hydrogen is a weaker greenhouse gas than methane, it would be ideal if at least a portion of hydrogen gas emissions could also be reduced. For this purpose, it would be desirable to combine or remove at least a portion of the hydrogen gas produced in the lumen. Phloroglucinol degradation in the lumen has been found to promote the sequestering of excess hydrogen that would otherwise be used for methane production (see Martinez-Fernandez G, et al., Front Microbiol. 2017 Oct 5;8:1871. doi:10.3389 / fmicb.2017.01871). Other compounds may also be used to promote the growth of microorganisms in the hydrogen-utilizing lumen, thereby reducing the partial pressure of hydrogen in the lumen and the amount of this gas exhaled by animals. Accordingly, in one embodiment, the bolus of the present disclosure preferably comprises a hydrogen sequestering agent selected from the group of hydrogen sequestering agents consisting of fumaric acid, sodium fumarate, phenolic compounds, phloroglucinol, gallic acid, resorcinol, catechol, hydroquinone, and pyrogallol.

[0199] The bolus described herein, specifically a bolus containing a hydrogen-sequenced compound, may in some embodiments also be a small bolus such as a pill, tablet, or pellet. Such a small bolus may be administered as is, or by adding it to an animal's feed, for example, in the form of a tablet, pill, or pellet, thereby the bolus being consumed by the animal. For example, such a small bolus may be added to the feed of an animal consuming it in such an amount that the amount of feed consumed contains about 20 g of the hydrogen-sequenced compound per 1 kg of dry matter feed. Here, the compound is contained in the mixed bolus. For example, an animal such as cattle consuming about 15 kg to 25 kg of dry matter feed per day may consume about 300 to 500 g of the hydrogen-sequenced compound contained in the bolus described herein per day, for example, as part of the pill, tablet, or pellet described herein.

[0200] In one embodiment, in any bolus described herein, the methane inhibitor is selected from the group consisting of bromoform, monensin, nisin, lecithin, lauric acid, myristic acid, linoleic acid, and phospholipids, and combinations thereof. Preferably, the phospholipid comprises one or more polyunsaturated fatty acids, more preferably, the phospholipid comprises one or more fatty acids selected from 18-carbon polyunsaturated fatty acids (C18 PUFAs), palmitic acid, stearic acid, and oleic acid. In one embodiment, the methane inhibitor is mixed with a carrier and contained in the bolus.

[0201] It is understood that the controlled release of methane inhibitors through a housing can be influenced by several factors. For example, controlled release may be influenced by the affinity of the methane inhibitor to the support contained in the bolus of the disclosure, in which the support may play a role in the diffusion of the methane inhibitor through the housing of the bolus containing the housing. It is understood that more polar support, or support containing a higher degree of polar functional groups, will have a higher affinity for polar inhibitors than less polar support, or support having a lower degree of functional groups.

[0202] The relative affinity of the compound forming the housing described herein (in the bolus containing the housing) and the core of the bolus to the methane inhibitor may also affect the controlled release of the methane inhibitor from the core. For example, having a housing with a relatively low affinity for the methane inhibitor compared to the affinity of the carrier to the methane inhibitor may be a factor in controlling the release rate of the methane inhibitor from the core.

[0203] In one embodiment, the methane inhibitor contained in the core of the bolus is a haloform, preferably selected from the list of bromoform, chloroform, iodoform, and combinations thereof. In a particularly preferred form, the haloform may be bromoform (CHBr3). Bromoform is reactive and has a short half-life in animals (0.8 h in rats, 1.2 hours in mice; US Dept of Health, 2003). It is a liquid at room temperature and is denser than water. Previous studies have not demonstrated significant increases in levels of residue in meat and tissue from slaughtered steers after a 48-hour withdrawal period (Kinley et al. Mitigating the carbon footprint and improving productivity of ruminant livestock agriculture using a red seaweed, Journal of Cleaner Production 259 (2020) 120836), nor in milk (Roque et al. Inclusion of Asparagopsis armata in lactating dairy cows' diet reduces enteric methane emission by over 50 percent; Journal of Cleaner Production 234 (2019) 132-138).

[0204] The use of bromoform can offer several advantages. For example, it has high potency at relatively small doses, which allows a single device to deliver a sufficient amount of methane inhibitor over an extended period. In addition, bromoform also has a relatively high density, which increases the overall weight of the bolus, allowing it to be retained in the lumen. That is, it sinks to the ventral side of the lumen rather than floating, reducing regurgitation. In one embodiment, the bolus may contain haloform, preferably bromoform, in an amount of 10% (by weight) to 80% (by weight), preferably 20% (by weight) to 50% (by weight).

[0205] Furthermore, the methane inhibitor may be synthetic or derived from naturally occurring sources, such as plants, including algae. In one embodiment, the methane inhibitor is Asparagopsis or a derivative thereof. The methane inhibitor can be obtained from Asparagopsis, for example, by extraction. For example, algal lysis can be achieved by breaking the cell wall or membrane of the algae and separating the methane inhibitor from the remaining algal biomass. Algae such as Asparagopsis, or parts thereof and derivatives thereof, may be included directly or in a bolus as a source that releases a methane inhibitor such as bromoform.

[0206] In another embodiment, the methane inhibitor is monensin. Monensin is a polyether carboxylic acid ionophore that can modify rumen fermentation dynamics by selectively inhibiting the growth of Gram-positive bacteria that produce most of the acetic acid, lactic acid, and hydrogen in the rumen that can contribute to methane formation. Advantageously, monensin is also known to prevent ketosis in ruminants. Administration of a ketosis inhibitor may be beneficial when administering a methane inhibitor such as haloform. Another methane inhibitor that can be used in addition to or as an alternative to monensin is the bacteriocin nisin. Both nisin and monensin inhibit methanogenic bacteria primarily by increasing the permeability of their cell membranes.

[0207] Lecithin is well known and has an effect on rumen fermentation and digestion, and therefore may contribute to methane suppression. For example, soy lecithin may be suitable in this context.

[0208] Certain saturated and unsaturated fatty acids may also be used due to their ability to influence rumen fermentation and the microbial composition in the rumen, and therefore to affect the methane production potential in the rumen. In one embodiment, the methane inhibitor is selected from lauric acid, myristic acid, and linoleic acid.

[0209] Phospholipids are known to typically contain a glycerol molecule, the carbon atom of which is linked to two fatty acids and a phosphate group, where the fatty acids and phosphate group are attached to the glycerol molecule via ester bonds. In one embodiment, the bolus described herein comprises a methane inhibitor selected from the group consisting of phospholipids comprising a glycerol molecule linked to a phosphate group and two fatty acids via ester bonds, preferably wherein the phospholipid comprises one or more polyunsaturated fatty acids, more preferably wherein the phospholipid comprises one or more fatty acids selected from 18-carbon polyunsaturated fatty acids (C18 PUFAs), palmitic acid, stearic acid, and oleic acid.

[0210] Considering the phospholipids that can be suitably used, phospholipids containing one or more fatty acids having a methane-relaxing effect are preferred. For example, saturated fatty acids (SFAs) including, for example, laurin (C12), myristicin (C14), or palmitin (C16), and stearic acid (C18) are known to suppress methane production in the rumen. Furthermore, polyunsaturated fatty acids (PUFAs) such as C12 and C18 PUFAs strongly oppose methane production. Without attempting to limit ourselves to this presumed effect of these fatty acids, the presence of these fatty acids in the rumen is now thought to have an impact on the microbiome in the intestinal tract of ruminants. Specifically, such fatty acids are thought to be able to act against rumen methanogens that colonize the rumen and capture H2 and CO2 that would otherwise be produced by other fermentation members of the rumen microbiome and produce methane (CH4). In a preferred embodiment, the phospholipid is selected from lecithin, phosphatidylcholine, and derivatives of the compounds described above.

[0211] In one embodiment, at least 50 wt% of the bolus of the disclosure contains a methane inhibitor. In another embodiment, at least 60 wt% and at least 70 wt% of the bolus of the disclosure contains a methane inhibitor.

[0212] It should be understood that the ratio of methane inhibitor to carrier or to total bolus weight may be selected to optimize the function of the bolus, for example, to suit a desired emission profile for each inhibitor(s). In one embodiment, a haloform, preferably bromoform, is included in the core of the bolus of this disclosure in an amount between 10 wt% and 80 wt%, preferably between 15 wt% and 70 wt%. In the context of methane inhibitors such as haloform or bromoform included in the bolus described herein, the term “wt%” as used herein means the weight percentage of the methane inhibitor based on the total weight of the bolus. In relation to the above methane inhibitors, suitable carrier materials may be advantageously used in the bolus of this disclosure having a high ability to retain the methane inhibitor. Due to the volatility of some methane inhibitors, such as bromoform, and their reactivity with many compounds, including organic compounds, it is inherently difficult to include such methane inhibitors in a stable manner and at high concentrations (so as to reduce the size of the formulation) within delivery devices such as boluses. One example of a particularly suitable carrier material is fumed silica, preferably hydrophobic fumed silica, which may consist of amorphous silica particles that can be fused into branched particles. Fumed silica, for example, available as a powder, offers low bulk density and high surface area. Using fumed silica as a carrier in a bolus will stabilize the formulation, improve its stability, and increase the loading capacity of the drug formulation for methane inhibitors, specifically haloforms such as bromoform. Therefore, when bromoform is used as a methane inhibitor, it may be preferable for the bolus to contain fumed silica.

[0213] For some methane inhibitors, it may be preferable to incorporate one or more openings into the housing of the bolus to facilitate the release of the methane inhibitor from the bolus. In a further aspect, the present disclosure relates to a bolus for administration to ruminants, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core comprising a methane inhibitor and a carrier; and a housing comprising the core, wherein the methane inhibitor is selected from the group consisting of monensin, lauric acid, myristic acid, and linoleic acid; and the housing comprises at least one opening that exposes the core to the environment surrounding the bolus. Preferably, the housing material comprises PLA and PBAT.

[0214] This embodiment may be particularly useful in cases where compounds other than haloform are used as methane suppressants.

[0215] In one embodiment, a bolus according to any aspect or aspect includes a housing, wherein the housing includes a stabilizer. In a preferred embodiment, the stabilizer is selected from the group consisting of surfactants, plasticizers, phthalates, and triglycerides. In a more preferred embodiment, the stabilizer is selected from the group consisting of lecithin, nitriles, and triacetin.

[0216] For example, stabilizers can have an effect on the stability of a housing by introducing more flexibility into the housing material or material blend, or by reducing the brittleness or tendency of the housing material or material blend to become brittle. In the case of a housing containing a blend of different materials, stabilizers can also be beneficial by promoting the mixing efficiency of the blend components. This produces a more homogeneous housing, which may be more stable against the forces acting through the digestive system of ruminants. For example, lecithin is known to be a suitable surfactant and stabilizer used in food and pharmaceuticals. The average person skilled in the art will know of further compounds that can be suitably used as stabilizers forming part of a housing.

[0217] In one embodiment, the core contained within the bolus comprises at least one filler. This embodiment relates to any bolus described herein. The use of a filler can provide additional internal material to the bolus and provide sufficient internal stability to counteract forces acting on the bolus from the outside. In addition, the filler can provide additional filler material to uniformly distribute a methane inhibitor within it (inside the bolus) while being well tolerable to ruminants at the same time, without promoting any significant undesirable interactions.

[0218] In a preferred embodiment, at least one filler is a stabilizer. In another preferred embodiment, at least one filler is selected from the group consisting of gelatin, milk, milk derivatives, infant formula, powdered milk, triglycerides, medium-chain triglycerides and their oils, ethanol, lecithin, tween, xanthan gum, cellulose derivatives, alkylcellulose, ethylcellulose, hydroxypropyl methylcellulose (HPMC), zein, and surfactants.

[0219] In another preferred embodiment, the core contained in any bolus described herein comprises gelatin. In yet another preferred embodiment, the core contained in any bolus described herein comprises milk, milk derivatives, infant formula, or powdered milk. In yet another preferred embodiment, the core contained in any bolus described herein comprises powdered milk. In yet another preferred embodiment, the core contained in any bolus described herein comprises at least one stabilizing and / or filling protein. In yet another preferred embodiment, the core contained in any bolus described herein comprises casein and / or zein. In yet another preferred embodiment, the core contained in any bolus described herein comprises a cellulose derivative, preferably alkylcellulose, ethylcellulose, and / or hydroxypropyl methylcellulose (HPMC). In yet another preferred embodiment, the core contained in any bolus described herein comprises a cellulose derivative, preferably alkylcellulose, ethylcellulose, hydroxypropyl methylcellulose (HPMC), combinations thereof, and copolymers thereof.

[0220] In one embodiment, the core contained in the bolus according to any aspect or embodiment comprises at least one PEG glyceride composed of mono, di, and triglycerides and mono and diesters of PEG, preferably comprising PEG esters of palmitin, stearin, and / or lauric acid.

[0221] In one embodiment, the core contained in a bolus according to any aspect or embodiment described herein comprises at least one surfactant, preferably a nonionic water-dispersible surfactant. In a preferred embodiment, the surfactant comprises a mixture of glycerides and fatty acid esters, preferably a mixture of mono, di, and triglycerides and PEG.

[0222] In another embodiment, the surfactant comprises a polyoxylglyceride, more preferably an oleoyl polyoxyl-6 glyceride. In one embodiment, the surfactant comprises mono, di, and triglycerides of oleic acid (C18:1) and PEG-6 (MW300) mono and diesters.

[0223] In another embodiment, the surfactant includes mono and diesters of caprylic (C8) and capric (C10) acids.

[0224] In another embodiment, the surfactant comprises mono, di, and triglycerides of lauric (C12) and stearic (C18) acids, and PEG-6 (MW300) mono and diesters.

[0225] In another embodiment, the surfactants include mono, di, and triglycerides of lauric acid (C12), as well as mono and diesters of PEG-32 (MW1500).

[0226] In one embodiment, the bolus core comprises zein. Zein is a composition comprising at least one protein and a prolamin, which can typically be found in protein bodies, for example, in the endosperm of a corn kernel. Due to the amphiphilic nature of the protein, zein is also useful in forming a protective coating. The product can be encapsulated based on the ability of zein to associate or self-assemble when the solution polarity changes toward a more hydrophilic environment. Thus, zein can be used not only as a core component of the bolus of this disclosure, but also suitably applied as a coating of the bolus.

[0227] The bolus of this disclosure may also be used, for example, as a delivery platform to allow the local sustained release of other methane-inhibiting and non-anti-methane-producing molecules. In one embodiment, in addition to or instead of the methane inhibitor, the bolus of this disclosure comprises an active agent, wherein the active agent is not a methane inhibitor. In one embodiment, in addition to the methane inhibitor, the bolus of this disclosure comprises an active agent, wherein the active agent is not a methane inhibitor. In one embodiment, instead of the methane inhibitor, the bolus of this disclosure comprises an active agent, wherein the active agent is not a methane inhibitor.

[0228] In one embodiment, in addition to or instead of the methane inhibitor, the bolus of the present disclosure comprises an active agent, wherein the active agent is not a methane inhibitor, and wherein the release of the active agent is local and / or sustained release in the intestinal system of a ruminant. Preferably, the release of the active agent is local and sustained release in the intestinal system of a ruminant.

[0229] In one embodiment, the active agent, which is not a methane inhibitor, is selected from the group consisting of anti-inflammatory agents, analgesics, and anthelmintics. However, the active agent may be any health and / or growth-promoting and / or sustained-release agent known in the art, or a combination of such agents. For example, the anti-inflammatory agent and / or analgesic may be selected from nonsteroidal anti-inflammatory drugs (NSAIDs). In one embodiment, the active agent is selected from meloxicam and ketoprofen. For example, the anthelmintic may be albendazole. Therefore, in one embodiment, the active agent is albendazole. Further active agents may also be antibiotics selected from the group consisting of penicillin, tetracycline, macrolides, monensin, ceftiofur, florfenicol, tilmicosin, enrofloxacin, and tulathromycin. Therefore, in a preferred embodiment, the active agent is selected from the group consisting of anti-inflammatory agents, analgesics, antibiotics, and anthelmintics. In a more preferred embodiment, the active agent is selected from the group consisting of meloxicam, ketoprofen, penicillin, tetracycline, macrolide, monensin, ceftiofur, florfenicol, tilmicosin, enrofloxacin, tulathromycin, and albendazole. Other compounds routinely and commonly administered to ruminants, and substances having one or more beneficial effects on ruminants, are known in the art, and an average person skilled in the art knows how to suitably implement the substances relating to the bolus of this disclosure. Such compounds also include, but are not limited to, growth promoters, milk secretion promoters, and sustained-life enhancers. Sustained-life enhancers are agents that improve the sustained-life of ruminant rearing. This improvement may be a reduction in pollution, for example, a reduction in greenhouse gas emissions.

[0230] In one embodiment, the core of the bolus described herein may contain one or more metal particles (preferably steel particles), where the particles are preferably round. For example, the total mass of all particles per bolus may be at least 100 g. Incorporating metal particles into the bolus adds weight to the bolus, allowing it to be more effectively held in the rumen of an animal and preventing regurgitation of the bolus after administration.

[0231] The bolus of this disclosure may be used in the treatment of animals. In one aspect, this disclosure relates to the bolus of this disclosure for use in the treatment of ruminants. In a preferred embodiment, this disclosure relates to the bolus of this disclosure for use in the treatment of cattle or sheep.

[0232] In another aspect, the Disclosure relates to a bolus of the Disclosure for use in reducing methane emissions in ruminants. In a preferred embodiment, the ruminant is a cattle or a sheep. In a further embodiment, the Disclosure provides a method for administering a methane inhibitor to an animal, the method comprising the step of administering the bolus of the Disclosure to the animal. In a further embodiment, the Disclosure provides a method for reducing methane production in the rumen of a ruminant, the method comprising the step of administering the bolus of the Disclosure to the ruminant.

[0233] In a preferred embodiment, the bolus may be configured to be administered to ruminants, which may include beef or dairy cattle, sheep, goats, buffalo, deer, moose, giraffes, or camels.

[0234] In one embodiment, the bolus may be adapted to reduce the emission of one or more greenhouse gases ("GHGs") from ruminants.

[0235] In one embodiment, the ruminant could also be a goat or a deer.

[0236] The reduced methane emissions and / or greenhouse gas emissions from ruminants are expected to be lower compared to ruminants not treated with the bolus or methane inhibitor of this disclosure. In one embodiment, methane emissions in ruminants may be reduced by at least 30%, preferably at least 50%, more preferably at least 70%, 80%, and most preferably at least 90%. In another embodiment, methane emissions in ruminants may be reduced by at least 99%. In a preferred embodiment, the bolus is administered orally. In another preferred embodiment, the bolus is configured to remain in the rumen after administration.

[0237] The bolus is delivered orally into the rumen of the ruminant to be treated and enters the rumen via the esophagus. In the rumen, gastric juices (and other substances such as plant fibers) may act to erode or dissolve the core so that it releases methane inhibitors over time. Alternatively, the contents of the bolus, such as methane inhibitors, may diffuse or leach out of the bolus into the rumen.

[0238] If the bolus includes a housing, the housing remains substantially intact throughout the duration of the procedure. In cases where the bolus housing includes one or more openings, these openings allow gastric fluid and possibly fibrous material to come into contact with the core.

[0239] The core and housing may be designed to facilitate the release of the methane inhibitor over the period during which the animal is to be treated. The bolus may be adapted to release the methane inhibitor over a period of at least 3 months, preferably at least 6 months, more preferably 12 months, and potentially up to 2 years. Preferably, the release rate of the methane inhibitor may be calculated based on the weight of the ruminant to be treated and the type of inhibitor used. It will be understood that the desired release rate may vary among animals. Typically, the desired release rate may be calculated from the amount of inhibitor / weight of the animal. Alternatively, the desired release rate may also be calculated based on the amount of feed consumed by the animal. Particularly preferred release rates for bromoform as an exemplary methane inhibitor include approximately 0.1 to approximately 0.5 g / day, more preferably approximately 0.2 g / day.

[0240] Ruminants may also be treated by multiple boluses according to this disclosure to achieve a preferred dose of the methane inhibitor. This may allow for the preparation of boluses having a certain concentration and total load of the methane inhibitor. Multiple of these boluses may be administered to the animal simultaneously or sequentially. This will allow the desired dose to be provided to the animal. This may be particularly beneficial to allow the use of boluses in animals requiring different doses of the inhibitor, for example, larger or smaller animals, or to compensate for natural growth over time.

[0241] Ruminants may also be treated with multiple boluses according to the present disclosure to achieve a preferred combination of substances, where different substances may be administered simultaneously. For example, a combination of boluses may be preferred in which one bolus provides a methane inhibitor and further boluses provide different active agents such as antibiotics or other health-improving compounds for ruminants.

[0242] A bolus can be adapted to deliver a certain dose of an inhibitor directly into the animal's rumen. For example, bromoform can be released at a rate that effectively reduces or eliminates methane production during digestion. This would reduce greenhouse gas emissions, particularly methane, from animals, and thus reduce the environmental impact of agriculture.

[0243] It will be understood by those skilled in the art that the size, thickness, and / or dimensions of the bolus, including the core and, if provided, the housing, can be adjusted depending on the dose of inhibitor to be delivered to the ruminant without departing from the spirit and scope of this disclosure. For example, smaller sized bolus may be adapted for use in smaller ruminants such as sheep or goats, and larger bolus may be used for larger ruminants such as cattle. Preferably, the bolus has a weight of less than 180 g. For example, each cattle may be administered two bolus, each having dimensions of about 75 mm in length and about 34 mm in width, thereby each of these bolus may have a weight of about 80 g.

[0244] In addition, reducing methane production can provide benefits to animal production. Bolas can improve the conversion of ruminant feed for animal production. For example, by reducing methane production during digestion, this could lead to more efficient utilization of ingested feed, resulting in improved growth and weight gain, or other production such as milk or meat production. As a result, farmers may be able to improve efficiency by either ensuring greater productivity for a given feed volume or by reducing feed accordingly. In addition, the synergistic effects arising from the composition for the core, as well as the combination of carriers and inhibitors (one or more), may enable the provision of long-term delivery devices for slow-release to improve animal productivity and / or reduce greenhouse gas emissions.

[0245] In further aspect, the disclosure relates to a method for manufacturing a bolus as defined herein. The manufacturing method may include the following steps: (1) providing a housing, preferably made of a polymer material, more preferably a biodegradable polymer, or preferably a housing of a material disclosed herein; and (2) filling the housing with a core preferably comprising a material disclosed herein. Here, the bolus includes: a core comprising a methane inhibitor and a carrier that inhibits methane production in the rumen of a ruminant, and a housing that houses the core. Providing the housing in step (1) may be done, for example, by 3D printing or injection molding, without limitation. Filling the housing with the core in step (2) may be done, for example, by melting or mashing and mixing the core material, and filling the housing with core material components or mixtures while the components are fluid or at least flexible or malleable.

[0246] Preferably, the method for manufacturing the bolus may further include the step (3) of sealing the housing containing the core with a cap. The housing may be sealed with a cap by friction welding the cap to the housing. This is advantageous compared to screw-on or glued caps, which may loosen or be pushed out of the housing when the bolus is exposed to chemical and mechanical stresses and turbulent motion in the animal's lumen.

[0247] Providing the housing in step (1) can occur using any technique that should be known to those skilled in the art. For example, a suitable material can be extruded into a desired shape defining the cavity. Alternatively, additive manufacturing processes or molding processes can also be used to construct the housing shape defining the cavity. For example, injection molding, 3D printing, or hot melt extrusion processes.

[0248] The core filling step (2) may include, for example, one or more of the following steps: melting and / or mashing a carrier material to provide a melted and / or mashed carrier material; adding one or more methane inhibitors to the melted and / or mashed carrier material; mixing the methane inhibitors and the melted and / or mashed carrier material to produce a substantially homogeneous mixture; and filling the prepared housing with the substantially homogeneous mixture.

[0249] For boluses without a housing, step (1) of forming the housing may be omitted, and step (2) may be a step of forming the core. This may include, for example, one or more of the following steps: melting and / or mashing the carrier material to provide a melted and / or mashed carrier material; adding one or more methane inhibitors to the melted and / or mashed carrier material; mixing the methane inhibitors and the melted and / or mashed carrier material to create a substantially homogeneous mixture; and forming the substantially homogeneous mixture into a desired shape. It should be understood that the step of forming the substantially homogeneous mixture into a desired shape may involve providing the mixture to a mold.

[0250] It should be understood that a substantially homogeneous mixture contains one or more methane inhibitors at concentrations sufficient to achieve the desired release profile for the methane inhibitor upon administration of the device to ruminants. The concentrations may vary depending on the type of ruminant to be treated, the shape and dimensions of the device, or the desired release profile to be achieved.

[0251] The method also includes a step that allows a substantially homogeneous mixture to cool, especially if it has been previously heated and melted. As it cools, the carrier material hardens and takes shape according to the shape of the mold or housing provided inside.

[0252] This disclosure also provides boluses that can or can be obtained by carrying out the method of manufacturing boluses described herein.

[0253] In another aspect, the present disclosure relates to a methane inhibitor for use in reducing methane emissions from ruminants, wherein the methane inhibitor is selected from the group consisting of bromoform, monensin, lecithin, lauric acid, myristic acid, linoleic acid, and phospholipids, preferably wherein the phospholipid comprises one or more polyunsaturated fatty acids, more preferably wherein the phospholipid comprises one or more fatty acids selected from 18-carbon polyunsaturated fatty acids (C18 PUFAs), palmitic acid, stearic acid, and oleic acid. In a preferred embodiment, the ruminant is a cattle.

[0254] In another aspect, the present disclosure relates to a method for treating ruminants to reduce methane emissions from said ruminants, comprising administering to said animals a methane inhibitor selected from the group consisting of bromoform, monensin, lecithin, lauric acid, myristic acid, linoleic acid, and phospholipids, preferably wherein the phospholipid comprises one or more polyunsaturated fatty acids, and more preferably wherein the phospholipid comprises one or more fatty acids selected from 18-carbon polyunsaturated fatty acids (C18 PUFAs), palmitic acid, stearic acid, and oleic acid. In a preferred embodiment, the ruminant is a cattle. The inventors believe that the techniques described herein may offer several benefits. These benefits may be the result of unique synergistic interactions between different aspects and embodiments of the techniques. Thus, the techniques of the present disclosure are described based on the inventors' current understanding of such possible interactions. It should be understood that any aspect or embodiment described herein, or the interaction of two or more aspects / embodiments, may form a separate disclosure. Modified release formulation

[0255] A modified release formulation is a formulation that alters the timing, rate, or site of release of the active ingredient in order to achieve clinical outcomes that are not achievable with the unmodified release formulation.

[0256] One form of modified release is temperature-dependent release, where the release of the active ingredient changes in response to temperature differences. This allows for release only under favorable temperatures and potentially enables control over the release site. While temperature-dependent release offers advantages in administration, it also offers advantages in storage, potentially allowing storage at harsher temperatures, which is particularly beneficial in the harsher storage conditions often encountered when handling livestock.

[0257] Another form of modified release is sustained release, prolonged release, or sustained release, which slows down the release of the active ingredient so that one dosage form can provide release of the active ingredient over a longer period of time. This has the advantage of reducing the frequency of administration. Since once-daily oral administration is generally considered acceptable, reducing the frequency of administration in humans is usually only a few hours. The benefits of sustained-release dosage forms are even more pronounced in the treatment of livestock. Daily or even weekly administration can be contraindicated for many livestock, especially those that need to be gathered for treatment. Treatment may only be feasible by weekly, bi-weekly, monthly, every six weeks, every eight weeks, every two months, or every ten weeks. This represents an extraordinary extension of the release of the dosage form to be achieved.

[0258] Slow release can be primarily due to the core. Slow release can be primarily due to the housing. The amount of extension can vary from small to large, for example, from one day to several months or even one year, depending on the composition of the core and housing.

[0259] Sustained release may be a release profile in which the bolus releases the methane inhibitor at at least 20%, at least 30%, at least 40%, or at least 50% of the maximum release rate 24 hours after the maximum release rate is achieved. Sustained release may be a release profile in which the bolus releases the methane inhibitor at at least 20%, at least 30%, at least 40%, or at least 50% of the maximum release rate one week after the maximum release rate is achieved. Sustained release may be a release profile in which the bolus releases the methane inhibitor at at least 20%, at least 30%, at least 40%, or at least 50% of the maximum release rate one month after the maximum release rate is achieved. Sustained release may be a release profile in which the bolus releases the methane inhibitor at at least 20%, at least 30%, at least 40%, or at least 50% of the maximum release rate six months after the maximum release rate is achieved.

[0260] Sustained release may be a release profile in which the bolus releases the methane inhibitor at at least 20%, at least 30%, at least 40%, or at least 50% of the maximum release rate 24 hours before the maximum release rate is achieved. Sustained release may be a release profile in which the bolus releases the methane inhibitor at at least 20%, at least 30%, at least 40%, or at least 50% of the maximum release rate one week before the maximum release rate is achieved. Sustained release may be a release profile in which the bolus releases the methane inhibitor at at least 20%, at least 30%, at least 40%, or at least 50% of the maximum release rate one month before the maximum release rate is achieved. Sustained release may be a release profile in which the bolus releases the methane inhibitor at at least 20%, at least 30%, at least 40%, or at least 50% of the maximum release rate six months before the maximum release rate is achieved.

[0261] Matrix systems, sustained release coatings, and other systems, for example sustained release particles within a matrix, may be used to prolong the release of the active ingredient. The housing of the present disclosure functions as a sustained release coating. The core of the present disclosure also has sustained release properties. In a preferred embodiment, the core of the present disclosure is a matrix system in which the active ingredient is homogeneously mixed into a core excipient (optionally as particles or granules with a non-sustained release carrier). Alternatives contemplated by the present disclosure include sustained release particles containing the active ingredient disbursed within a binder, wherein the binder may or may not have additional sustained release properties. Preferably, the active ingredient is dispersed directly and homogeneously into the other components of the core without excipients.

[0262] In a preferred embodiment, the matrix system in the core is hydrophobic (or water-insoluble with minimal swelling). Optionally, the core is a blend of hydrophobic and hydrophilic components. However, the release profile is largely controlled by the hydrophobic component.

[0263] In a preferred embodiment, the housing is hydrophobic. In some embodiments, the core is hydrophobic. Optionally, both the core and the housing are hydrophobic.

[0264] In any embodiment of any aspect of the present disclosure, the core comprises a methane inhibitor and a carrier, and the methane inhibitor is optionally dispersed in the carrier. Zero-order release

[0265] Zero-order release of an active ingredient, which is constant release of the active ingredient over the duration of release, is the goal of preferred embodiments of the dosage form of the present disclosure. Active ingredient

[0266] In some embodiments, the methane inhibitor is a haloform, including mixed haloforms. Preferably, the haloform is selected from chloroform, bromoform, iodoform, or combinations thereof; more preferably bromoform.

[0267] Optionally, the active ingredient or haloform may be approximately 20-90%, 30-80%, 40-80%, 50-70%, or 60% w / w of the core. Hydrophobic

[0268] In some embodiments, one or more of the housing and core are hydrophobic. Those skilled in the art will understand that hydrophobicity can be measured by analyzing the water contact angle using a goniometer. In some embodiments, the difference in static water contact angle between one or more of the housing and core is at least about 60°, at least about 50°, at least about 40°, at least about 30°, at least about 20°, at least about 15°, at least about 10°, or at least about 5° at 20°C. Ruminants

[0269] In some embodiments, the bolus is for or suitable for administration to the rumen of a ruminant. In some embodiments, the ruminant is a cattle, sheep, goat, or deer. In some embodiments, the ruminant is a cattle. In some embodiments, the ruminant is a sheep. Bolas

[0270] Broadly speaking, a bolus is a dosage form of a substance such as a drug, supplement, or metabolic modifier that has an individualized dose. In the context of this disclosure, a bolus may be a solid, semi-solid, or a combination thereof. A bolus may also be a combination of a liquid with a solid, semi-solid, or a combination thereof, provided that the liquid is encapsulated within the solid, semi-solid, or a combination thereof. A semi-solid may be a blend of a liquid with a solid or semi-solid substance. Bolus are typically used for oral administration to the gastrointestinal tract of animals, preferably to the rumen of ruminants. Bolus are swallowed, but may be administered with the assistance of a bolus gun or ball gun. Several versions of these are commercially available. Bolus may vary in shape, but are commonly round, elongated, or capsule-shaped. Bolus may vary in size to suit administration to the applicable animal. Bolus may be rigid or have a softer, more malleable consistency. A bolus can be in the form of a pill, capsule, or tablet, as long as the pill, capsule, or tablet can be administered using a bolus or ball gun, rather than being a smaller pill, capsule, or tablet of a size suitable for inclusion in animal feed.

[0271] In a preferred embodiment illustrated in Figure 22, the bolus (100) of the present disclosure comprises a housing (101) that encapsulates or substantially encapsulates a core (102). The housing further includes a sealed region (103). The housing is optionally about 0.5 to about 2.0 mm thick, about 0.8 to about 2.0 mm thick, about 0.8 to about 1.8 mm thick, about 0.9 to about 1.8 mm thick, about 1.0 to about 1.8 mm thick, about 0.8 to about 1.5 mm thick, about 0.9 to about 1.5 mm thick, about 1.0 to about 1.5 mm thick, or about 1.2 mm thick. The housing is optionally about 5 to about 15%, about 6 to about 12%, about 6 to about 10%, about 7 to about 15%, about 7 to about 12%, about 7 to about 10%, or about 8% w / w of the bolus. The core is optionally approximately 20–55%, 25–50%, 30–45%, 35–40% w / w, or 37% w / w of the bolus. The bolus further contains a densifying agent (104), which separates the sealed region of the housing from the core. The densifying agent is optionally approximately 30–75%, 40–70%, 45–65%, 50–60%, or 55% w / w of the bolus.

[0272] In some embodiments, the bolus comprises a therapeutically effective amount of a methane inhibitor and the following: • Approximately 5% to 15% (w / w) housing; • Approximately 20 to 55 (w / w) cores; and • Approximately 30 to 75 (w / w) high-density agent. Release and duration

[0273] In some embodiments, the bolus of the present disclosure comprises the methane inhibitor bromoform and is adapted to reach a maximum release rate of approximately 0.1 to approximately 0.5 g per day, more preferably approximately 0.2 g per day. Such a release rate can provide a sustained release of a haloform such as bromoform. Bolus having such a release rate is suitable for use in livestock such as cattle and sheep.

[0274] In some embodiments, the bolus may be adapted to exhibit an release rate into the rumen between 0.02 g and 2 g per day, preferably approximately 0.1 to 0.5 g of bromoform per day. When the bolus exhibits such a release rate for a methane inhibitor (e.g., a haloform such as bromoform), this can reduce methane production. The release rate of the methane inhibitor into the rumen may increase over time; that is, the release rate starts from zero upon administration to the animal and increases to a maximum due to several factors. However, the foregoing should not be seen as limiting, and other release rates are assumed to be within the scope of this disclosure.

[0275] In some embodiments, the bolus is formulated to deliver haloform to the rumen of a ruminant for at least approximately 8 weeks after administration. In some embodiments, the bolus is formulated to deliver haloform to the rumen of a ruminant for at least approximately 20 weeks after administration. Retention in the lumen

[0276] The length of time a bolus is retained in the lumen can be increased by formulating the bolus to have a much higher density than that of the fluid in the lumen. One way to achieve this outcome is to incorporate a densifying agent into the bolus. In some embodiments, the bolus further contains a densifying agent. The densifying agent is a component that increases the density of the bolus. The densifying agent may be a metal powder such as ZnO, a metal ball such as a steel ball, or other high-density material suitable for inclusion in the bolus. Preferably, the densifying agent increases the density of the bolus to 1.0 g / cm³. 3To increase the density to a much greater extent. In some embodiments, the densifier is a densifier matrix comprising a densifier and at least one veterinarily acceptable excipient. Optionally, the densifier matrix comprises a matrix material that is more hydrophobic than at least one carrier. Optionally, the densifier matrix comprises a wax. Optionally, the densifier is either dispersed in the core, in or on the housing, or separate from the core and housing (preferably in the housing). Optionally, the densifier is about 30 to about 75% w / w of the bolus, preferably about 45 to about 65%, or about 55% w / w of the bolus.

[0277] An alternative approach to increasing the length of time the bolus spends in the lumen is to ensure that the bolus is too large to pass out of the lumen while it is in it. This type of device is known to those skilled in the art and often involves components that remain in close contact with the bolus during administration and then expand to increase the size of the bolus after administration. These components have an area of ​​at least 4 cm² at at least one point. 2 , at least 5cm 2 , at least 6cm 2 This may include attaching further components to the bolus to increase its cross-section. The center of the cross-section does not need to be solid. Diffusion test

[0278] Diffusion testing is a common technique for evaluating the properties of a dosage form in vitro. The results of diffusion testing often correlate with the in vivo performance of the dosage form and are used in quality control testing to ensure consistent manufacturing of the dosage form.

[0279] The diffusion of haloform from the bolus to the surrounding solution of the present disclosure was tested without stirring over a period of several months in 1 L of 0.02 M phosphate buffer at pH 6.5 (simulating lumen pH) and 39°C (simulating lumen temperature). Samples of the buffer were taken daily and analyzed for tribromomethane by GC-FID.

[0280] Diffusion tests have similarity to dissolution tests. Dissolution testing involves placing a dosage form in a liquid of a specific pH and temperature with specific stirring, and determining the time it takes for the active ingredient to be released from the dosage form. The United States and European Pharmacopoeia (USP & EP) have standardized dissolution tests, see, for example, Chapter <711> of USP. However, due at least to the extended bolus length and size, these dissolution tests are not suitable for measuring diffusion of the dosage forms of the present disclosure. Method of Administration

[0281] In another aspect, the present disclosure provides a method of administering a methane inhibitor to a ruminant, the method comprising administering a bolus according to the present disclosure into the rumen of the ruminant.

[0282] In another aspect, the present disclosure provides a method of reducing methane production in the rumen of a ruminant, the method comprising administering a bolus according to the present disclosure into the rumen of the ruminant.

[0283] In an embodiment of the administration method of the present disclosure, the bolus administers haloform to the rumen of the ruminant for at least about 8 weeks after administration. Optionally, the bolus administers haloform to the rumen of the ruminant for at least about 20 weeks after administration.

[0284] In any embodiment of the present invention that refers to a bolus for release of a methane inhibitor to an animal, the animal is preferably a ruminant, and the bolus is preferably for release of the methane inhibitor into the rumen of the ruminant.

[0285] In an embodiment of the administration method of the present disclosure, after administration of the bolus, the bolus sinks below the liquid surface or to the bottom of the rumen.

[0286] In an embodiment of the administration method of the present disclosure, the bolus remains in the rumen for at least about 8 weeks or at least about 20 weeks after administration.

[0287] In embodiments of the administration method of this disclosure, after the release of the active ingredient, the bolus is broken down in the rumen. Optionally, the bolus is broken down until the remnants are small enough to safely pass through ruminants.

[0288] In embodiments of the administration method of this disclosure, after administration of the bolus, methane excreted by ruminants is reduced by approximately 50%, 55%, 60%, 65%, 70%, 75%, and 80% per day in g. Optionally, this reduction occurs within approximately 5, 10, or 15 days after administration of the bolus. Optionally, the reduction lasts for approximately 4, 6, 8, 12, 16, or 20 weeks. Optionally, the reduction continues at approximately 40–90%, 40–70%, 40–50%, 60–90%, or 70–90% per day in g over approximately 4, 6, 8, 12, 16, or 20 weeks.

[0289] In embodiments of the administration method of this disclosure, a second bolus is administered to ruminants approximately 8 to 20 weeks, 12 to 20 weeks, 8 to 16 weeks, or 12 to 16 weeks after the initial dose. Optionally, further bolus doses occur periodically at these intervals. Optionally, this dosage regimen results in ongoing methane reduction of approximately 40 to 90%, 40 to 70%, 40 to 50%, 60 to 90%, or 70 to 90% / day in g.

[0290] In some embodiments of the method of the present disclosure, the bolus of the present disclosure comprises the methane inhibitor bromoform and reaches a maximum release rate of approximately 0.1 to approximately 0.5 g per day, more preferably approximately 0.2 g per day.

[0291] In some embodiments, the bolus exhibits an release rate into the lumen between 0.02 g and 2 g per day, preferably approximately 0.1 to 0.5 g of bromoform per day. The foregoing should not be seen as limiting, and other release rates are assumed to be within the scope of this disclosure.

[0292] In some embodiments, the bolus exhibits near-zero-order release kinetics. In some embodiments, the bolus exhibits near-zero-order release kinetics 2 months, 4 months, and / or 6 months after administration. Production method

[0293] In yet another aspect, this disclosure provides a method for making a bolus, the method comprising: Selecting the core and housing, Inserting the core into the housing, Optionally, sealing the housing to enclose or substantially enclose the core inside the housing: Here, the core contains a methane inhibitor. Optionally, the core and housing may be prepared prior to their selection.

[0294] In some embodiments, the housing is configured to increase its permeability to the methane inhibitor when the housing is exposed to the lumen of a living animal. Optionally, the bolus further contains a densifying agent (preferably within the housing) either dispersed in or on the housing, or separated from the core and housing.

[0295] In yet another aspect, this disclosure provides a method for making a bolus, where the bolus does not include a housing, and the method is: This includes forming a core from a methane inhibitor and a support.

[0296] Optionally, forming the core may include the use of a mold. Optionally, forming the core may include the use of a housing and subsequent removal of the housing.

[0297] These methods may be used to prepare bolus dosage forms in accordance with the present disclosure.

[0298] In some embodiments, inserting the core into the housing occurs prior to inserting the densifying agent into the housing. In some embodiments, inserting the densifying agent into the housing occurs prior to sealing the sealing region of the housing.

[0299] In some embodiments, sealing a sealing region includes sealing two sections of the housing together. Alternatively, sealing encompasses sealing a cap. For example, sealing a sealing region includes attaching a cap to the sealing region of the housing, or sealing a cap already attached to the housing on the core (optionally, attaching it to another part of the sealing region of the housing). In some embodiments, sealing is by sealing or suturing. Optionally, sealing encompasses soldering and / or spin welding.

[0300] In some embodiments, the sealing region includes means for sealing the housing, and the housing is sealed using the sealing means. Optionally, the means for sealing the housing is a cap.

[0301] In some embodiments, after sealing the sealing region, the sealed region is formed from previously separated portions of the housing that have been melted and / or soldered together.

[0302] In some embodiments, the densifying agent is above room temperature when it is inserted into the housing. In some embodiments, at least one component of the densifying agent and / or the densifying agent matrix is ​​liquid when it is inserted into the housing.

[0303] In some embodiments, the housing is prepared by injection molding.

[0304] In some embodiments, the densifying agent and / or densifying agent matrix are in direct contact with the core. In some embodiments, the densifying agent and / or densifying agent matrix are in direct contact with the sealed region. Preferably, the densifying agent is in direct contact with both the core and the sealed region. In some embodiments, the densifying agent and / or densifying agent matrix are not in direct contact with either or both the core and the sealed region (for example, further separating components may be present to prevent contact).

[0305] In some embodiments, the sealing region includes means for sealing the housing. In some embodiments, the means for sealing the housing is a cap. In some embodiments, the sealing region includes previously separated portions of the housing that are melted and / or soldered together. Packing materials

[0306] In a further aspect, materials comprising acrylate polymers are provided for use in packing the boluses of the present disclosure.

[0307] In some embodiments, the acrylate polymer is PMMA.

[0308] In some embodiments, PMMA is the only polymer that comes into contact with the bolus when it is packed into the packing.

[0309] Unless otherwise stated herein, any aspect disclosed herein may be freely combined with any other aspect disclosed herein.

[0310] It will be understood that the disclosures disclosed and defined herein extend to all alternative combinations of two or more individual features mentioned or evident from the text or drawings. All of these different combinations constitute various alternative aspects of this disclosure. [Examples]

[0311] Boluses for administration to ruminants are known. Therefore, they can be made as is well known in the art, for example, as described in WO2022124914, which is incorporated herein by reference. Below, improved production methods for the boluses of the present disclosure are described in examples that are not limited thereto. Considering these examples, it will be clear how alternative boluses of the present disclosure can also be made. Example 1

[0312] The following provides a general description of how to make a bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: a core containing the methane inhibitor; and a housing covering the outer surface of the bolus, wherein the housing is permeable to the methane inhibitor when exposed to a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C; and / or, wherein at least a portion of the housing is configured to form one or more openings when exposed to a temperature between 28°C and the temperature present in the rumen of a living animal, preferably between 28°C and 42°C, allowing the methane inhibitor to exit the bolus through the opening(s).

[0313] In the first step, the housing is made by 3D printing or injection molding. The housing can have various bottle-like shapes, preferably cylindrical. The housing is typically made from a biodegradable polymer, and the housing material includes, for example, PLA and PBAT. The housing may have a wall thickness of about 1.2 mm and dimensions of, for example, about 35 mm (diameter) × about 73 mm (length). Then, the housing is perforated, for example, by drilling openings or holes therein. The holes may have an average diameter of, for example, about 0.1 mm to about 1 mm. The diameter of the holes / openings can vary within a given housing.

[0314] Next, the holes / openings are sealed by filling them with a compound that melts between the temperature present in the rumen of a living animal and 28°C. Exemplary compounds and compound mixtures that can be used for this purpose are disclosed herein. Alternatively, the perforated bolus may also be wrapped in foil having the melting temperature mentioned.

[0315] Optionally, a portion of the internal volume of the housing may be filled with a densifying agent composition, for example, containing steel balls.

[0316] In the next step, tribromomethane is added to ethylcellulose and mixed until a homogeneous paste is obtained. HPMC is then added to the paste and mixed until a homogeneous dough is obtained. This dough (e.g., 60 grams) is then pressed into a prepared housing (which optionally contains a densifying agent composition).

[0317] In a further step, the housing is sealed by adding a cap that is spin-welded onto the housing to seal it. The steps for producing the bolus can also be carried out in any alternative order, for example, by perforating the housing after filling it with core material, and / or by sealing the openings on the housing in the final step.

[0318] This specification also provides boluses of the present disclosure that can be produced by carrying out the method steps outlined above. Example 2

[0319] The following provides a general description of how to prepare a bolus for administration to ruminants, wherein the bolus is configured to release a methane inhibitor in the animal, and the bolus comprises: A core comprising a methane inhibitor in microencapsulated particles, the microencapsulated particles being dispersed in a composition comprising a carrier and optionally also comprising a dispersant; and a housing covering at least a portion of the core.

[0320] Methods for producing microencapsulated substances are known, for example, from US6458118B1 or US7105158B1, both of which are incorporated herein by reference. In such systems, a small amount of a drug, e.g., 1 microgram, is encapsulated in an inert material, e.g., a stable polymer. Such an approach can be applied to encapsulate a methane inhibitor, e.g., bromoform. After encapsulation, the encapsulated haloform is then filled into a bolus of the present disclosure. This embodiment can also be produced by encapsulating a porous support, such as mesoporous silica, in the bolus. Example 3

[0321] A housing-free bolus can be made without a housing, for example, by following the steps in Example 1. Instead, the core material can be further densified by adding additional fillers such as fumed silica. The core is then compressed into, for example, pellets or similar. Example 4

[0322] The following provides a general description of a bolus for administration to ruminants, and how to make such a bolus, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: Cores comprising methane inhibitors and carriers; and Housing including the aforementioned core: Here, the methane inhibitor is selected from the group consisting of monensin, lauric acid, myristic acid, and linoleic acid; the housing includes at least one opening that exposes the core to the environment surrounding the bolus.

[0323] Such a bolus containing an alternative methane-suppressing compound can be produced, for example, by following the steps outlined above in Example 1, and substituting bromoform with one or more alternative methane-suppressing compounds.

[0324] In this embodiment, it is also possible not to fill the holes / openings of the perforated bolus. Example 5

[0325] A general description of how to prepare a bolus is provided below. Herein, in addition to or instead of the methane inhibitor, the bolus comprises an active agent, wherein the active agent is selected from the group consisting of anti-inflammatory agents, analgesics, antibiotics, and anthelmintics; more preferably, wherein the active agent is selected from the group consisting of meloxicam, ketoprofen, penicillin, tetracycline, macrolides, monensin, ceftiofur, florfenicol, tilmicosin, enrofloxacin, tulathromycin, and albendazole.

[0326] Such boluses can be prepared by substitution according to the steps outlined above in any of Examples 1-4, in which the methane inhibitor is replaced by an alternative activator. Example 6

[0327] The following provides a general description of how to use the bolus described herein in a method of treating an animal, which includes administering the bolus described herein to the animal, where the animal is preferably a ruminant such as a cattle.

[0328] A bolus as described herein, for example, any of the boluses in Examples 1 to 5, can be administered orally (esophageal) into the rumen of a ruminant animal such as a cattle. After administration of the bolus, the animal can be allowed to graze freely while the bolus remains in the rumen. Administering an active agent, such as a methane inhibitor, within a bolus that provides a sustained release of the active agent has the advantage that the active agent does not need to be re-administered very frequently.

[0329] Certain parameters may be evaluated to determine the effectiveness of the active agent released from the bolus, as well as the tolerability of the administered bolus and active agent, such as a methane inhibitor. This allows for improvements to the medication regimen for each active agent and bolus type used. For example, animal feed intake, such as dry matter intake, and animal body weight may be recorded as indicators of animal health and well-being.

[0330] Where a bolus should release methane or greenhouse gas suppressants, the reduction of these emissions can be quantified by measuring animal gas emissions, such as methane, hydrogen, and / or carbon dioxide emissions, in a respiratory chamber, for example, by using a 4900C continuous emissions analyzer and measuring emissions every 3 minutes over a 48-hour period.

[0331] When a bolus should release an antibiotic or anthelmintic, its effect on the microbiome or parasites can be assessed by taking a sample from the animal's intestine and evaluating it for the presence of the microbiome or parasites. Methods for assessing and quantifying the microbiome and parasites include microscopy and cell culture methods, bacterial antibiotic challenge, and molecular methods such as polymerase chain reaction (PCR).

[0332] The evaluation of the described animals may be repeated periodically during the evaluation period to verify the occurrence of efficacy over time. When using haloform (e.g., bromoform) as a methane suppressant, the bolus may be configured to release an amount of 0.4 mg of haloform per kg of ruminant per day. In one embodiment, larger ruminants on average size (i.e., weighing about 350-400 kg) may be administered haloform (e.g., bromoform) at a dose of 200 mg / d. Example 7

[0333] The effect of temperature on the release behavior of boluses was tested by preparing boluses and placing them in 1 L Schott bottles containing 0.02 M phosphate buffer as described below. These bottles were stored at room temperature (room temperature, 25°C), 30°C (±2°C), and 40°C (±2°C) for comparison. Bolus preparation

[0334] Bolas (1 and 2) with two different bromoform loads were prepared using the carrier / matrix compositions shown in Table 2. Briefly, in each case, bromoform (purity >95%, ethanol as stabilizer: 1-3%) was added to ethylcellulose (EC) (ethoxyl: 48-49.5%; chloride <0.05%; apparent viscosity 41-49 mPa.s) to form a viscous paste using a mortar and pestle. Those skilled in the art will understand that other mixing apparatus may also be used, especially on an industrial scale. To this mass, hydroxypropyl methylcellulose (HPMC) (methoxyl content 19-24%; hydroxypropyl content 7-12%; apparent viscosity 75,000-140,000 mPa.s) was added in small aliquots and then properly fixed to form a homogeneous mix. This process was repeated until all the HPMC had been added and a homogeneous dough / matrix was formed.

[0335] Once the bromoform / EC / HPMC matrix was prepared, approximately 60 g (±1 g to the exact measurement) of it was loaded into the body of the housing. A densifier (approximately 100 g, ±5 g to the exact measurement) was added on top of the matrix. The densifier was paraffin wax and loose stainless steel (SS) shots (0.1-0.5 mm diameter) in a ratio of approximately 15:1 (w / w). The densifier matrix was introduced as a molten paraffin wax / stainless steel shot mix poured directly onto the top of the bromoform / EC / HPMC matrix. The housing was filled to ensure that there were no or only small air gaps. As an alternative densifier option, the densifier matrix may be a pre-fabricated tablet of stainless steel shot / paraffin wax or any other preferably high-density material. Since the purpose of the densifier is to ensure that the bolus has sufficient density so that it sinks in the buffer and does not float, the methods described for the densifier should not be considered limiting factors for the range of densifiers. From the perspective of ruminant application, those skilled in the art will understand that sufficient density is desirable to achieve an effective bolus that, once administered to a ruminant, can retain itself in the lumen. After adding both the carrier / matrix and the densifying agent to the housing body, the cap was attached to the body by spin welding. Alternatively, a soldering iron may be used for attachment.

[0336] In this case, the housing was prepared using injection molding technology from a blend of 90% polylactic acid (PLA) (average molecular weight approximately 145,000 g / mol; D-lactic acid 1.2%) and 10% polybutylene adipate terephthalate (PBAT) (average molecular weight approximately 80,000 g / mol). Each bolus housing had a length of 73 mm, a diameter of 35 mm, and a thickness of 1.2 mm. Those skilled in the art will understand that other techniques are also suitable in addition to injection molding.

[0337] Table 2. Composition of support / matrix in each bolus. TBM = Tribromomethane (bromoform); EC = Ethylcellulose, HPMC = Hydroxypropylmethylcellulose. [Table 2]

[0338] DSC data for EC suggests that it operates in an amorphous manner. Release test

[0339] Once boluses were prepared, they were kept in 1 L Schott bottles containing 0.02 M phosphate buffer (pH=6.5) and stored without stirring at room temperature (RT, 25°C), 30°C (±2°C), and 40°C (±2°C). The buffer was replaced daily, with the exception of weekends. Nevertheless, in all cases, a minimum of four daily release data points were collected per week. Bromoform (TBM) was quantified using GC-FID (Shimadzu, Nexus GC-2030). Briefly, in each case, 10 mL of sample was collected in a 15 mL Falcon tube using a 10 mL autopipette. To this, 1 mL of ethyl acetate (analytical grade, Merck) was added to each Falcon tube as the extraction solvent for TBM. The Falcon tubes were capped, mixed well using a Vortex, and centrifuged at 4000 rpm for 15 minutes. 0.5 mL of ethyl acetate was recovered and loaded into a GC vial. 200 μl of sample was injected using an autosampler and analyzed in splitless mode over 20 minutes using a ZB5HT 30m capillary column with a temperature lamp of 30-300°C and a nitrogen gas flow of 5 mL / min. TBM had a retention time of approximately 5 minutes. The peak area was compared to a calibration standard prepared in ethyl acetate to determine the mass (mg) of TBM in solution and correlated to quantify the daily TBM release in 1 L buffered solution. result

[0340] Figures 1 and 2 show the release profiles of boluses (types 1 and 2; Table 2) when placed in a buffer at different temperatures. Temperature-dependent release was observed. In both cases, it was observed that boluses began releasing TBM earlier and at a higher rate when placed at higher temperatures than at lower temperatures. Nevertheless, surprisingly, a bolus releasing approximately 150 mg TBM per day at 40°C (Figure 1) was measured to begin significant release from day 14 at 30°C. However, a bolus releasing approximately 80 mg TBM / day at 40°C (Figure 2) was observed not to significantly release TBM for more than 35 days. This suggests that multiple factors are at work in addition to the role of temperature alone. It is well established that increasing the temperature of the system leads to higher thermal energy, which can often contribute to increased diffusion. Furthermore, even higher temperatures can make the polymer more permeable, contributing to an increased release rate. The permeability of polymers can be influenced by further factors such as their composition and degree of crystallinity.

[0341] Without being constrained by theory, it is suggested here, surprisingly, that the plasticizing effect of TBM (not only as an active ingredient but also as a solvent) on the housing polymer appears to play a significant role in the release behavior. The inventors observed that the solvent content of the housing material increases when exposed to bromoform / water. For example, the solvent content of the PLA / PBAT housing in Example 7 was found to increase at least threefold when the housing was exposed to a bromoform solution of approximately 100 mg / L for approximately 10 days. The solvent content was assessed using a thermogravimetric analyzer (TA instruments; model TGA55). Briefly, approximately 5 mg (accurately measured) of sample was heated from room temperature to 200°C at a heating rate of 20°C / min in an inert nitrogen atmosphere, and the change in weight was recorded. Furthermore, the inventors observed that the glass transition temperature of the housing may decrease with increasing solvent content. The glass transition temperature of the PLA / PBAT housing in Example 7 was observed to drop by approximately 10%, from approximately 55°C to approximately 50°C. The glass transition temperature was evaluated using a differential scanning calorimeter (TA instruments, model DSC250). Briefly, approximately 5 mg (accurately measured) of housing material was loaded into a Tzero aluminum pan and heated under modulated conditions. The sample was heated from 0°C to 195°C at a heating rate of 1°C / min, an amplitude of 0.16, and a modulation period of 60 s. Solvent sorption further potentially causes swelling and increases permeability. Swelling of PLA in the presence of organic solvents and water is well established (Udayakumar M et al. (2020) Polymers (Basel), May 6;12(5):1065).

[0342] Such changes in release behavior can potentially also be achieved through other changes, such as the crystallinity of the support and / or housing matrix. For example, the inventors used amorphous EC in Example 7. Changes in the temperature and plasticizing effect of TBM as a solvent can contribute to changes in crystallinity. This can lead to increased free TBM in the matrix that can break the bonding interaction between EC and TBM in the matrix and diffuse into the housing. Furthermore, the solid state of the housing itself can be altered to a higher crystallinity by annealing using dry heating or by the use of a solvent. Such changes in crystallinity can also be used to modulate the permeability or diffusion coefficient. For example, the release of TBM from a bolus with a housing crystallized (measured by DSC) by exposure to high temperature (approximately 80°C) over several hours showed reduced TBM release over a week compared to a bolus with a housing that was not treated with heat and therefore had a more amorphous content. Example 8

[0343] The effect of housing thickness on the emission profile was investigated using three different housing thicknesses: 0.90 mm, 1.2 mm, and 1.5 mm. The housing composition was the same 90% polylactic acid (PLA) and 10% polybutylene adipate terephthalate (PBAT) blend described in Example 7. Bolus preparation

[0344] The boluses were assembled by loading them with the matrix (bolus 2 matrix - Table 2) and the densifying agent matrix, and then capping them as described in Example 7. Release test

[0345] Each bolus was placed in 1 L of 0.02 M phosphate buffer as described in Example 7, and the amount of TBM released from the bolus daily was quantified. result

[0346] Bolus with a housing thickness of 0.9 mm began to release significantly from day 10 (Figure 3). Bolus with a housing thickness of 1.2 mm began to release significantly from day 29, and bolus with a housing thickness of 1.2 mm began to release significantly from day 35. The release rate was greater with thinner housings. This suggests an inverse relationship between housing thickness and release rate, and a direct relationship between housing thickness and TBM release lag period. Here, lag period refers to the time during which the bolus does not release TBM significantly. Example 9

[0347] The effects of buffer (water) or TBM on the housing material were tested. The bolus housing was the same as that described in Example 7. The housing was kept together with the bolus in 1 L of buffer at 40°C. Changes in the volatile content and glass transition temperature of the housing were evaluated at the end of 25 days. Volatile content was analyzed by heating an approximately 5 mg (accurately measured) sample from room temperature to 200°C at 20°C / min using a thermographic analyzer. The weight loss during the process was taken as the volatile content and expressed as a percentage relative to the original mass.

[0348] The glass transition of the material was evaluated during modulated differential scanning calorimetry (MDSC). Briefly, approximately 5 mg (accurately measured) of sample was loaded into a Tzero aluminum pan and heated from 0 to 195°C at a heating rate of 1°C per min with an amplitude of ±0.16 and a modulation period of 60 s. The glass transition temperature was taken from the reversible heat flow curve and analyzed using TRIOS software from TA instruments.

[0349] The test results are shown in Table 3. Exposure of PLA / PBAT to water / bromoform increases the total volatile content of the housing and lowers its glass transition temperature. The data suggest that the housing material has the ability to retain the solvent within it, and likely acts as a reservoir for bromoform itself, modulating its release. This also suggests that the presence of bromoform and water plasticizes the housing. Plasticization likely promotes further release of bromoform over time.

[0350] Table 3. Effects of buffering agent (water) or TBM on housing materials. TBM = Tribromomethane. [Table 3] Example 10

[0351] Surprisingly, TBM formed organogels with lecithin (purified, 98% acetone-insoluble, <1% residual water) and polymethyl methacrylate (PMMA) (molecular weight 450-550 kDa). With lecithin, the gel preferably >40% w / w The gel was formed by the TBM concentration. Here, the gel can also be formed at lower concentrations by using water. Water potentially contributes to increased hydrogen bonding in the matrix and stabilizes the 3D gel structure. In PMMA, the gel was preferably formed by a TBM concentration of >10% w / w.

[0352] Different boluses were prepared by loading lecithin and PMMA gels with different TBM concentrations, and their release profiles were evaluated. Bolus preparation

[0353] Lecithin / TBM gels were prepared by adding TBM to lecithin and mixing in a beaker. The lecithin dissolves and may form a yellowish, translucent, viscous mass at the beginning of mixing. This will later begin to gel. Those skilled in the art will understand that heat may be used. Three different gels with different bromoform loads of 45, 60, and 70% were prepared as outlined in Table 4.

[0354] PMMA / TBM gels were prepared by adding TBM to PMMA in a beaker. Those skilled in the art will understand that heat may be used. The mixture may dissolve and form a translucent, viscous mass. This will change into a gel upon prolonged standing. Two different gels were prepared with 45% and 70% TBM loads (Table 4).

[0355] Table 4. Composition of support / matrix in each bolus. TBM = Tribromomethane; PMMA = Polymethyl methacrylate. [Table 4]

[0356] The bolus housing was the same as that described in Example 7. As described in Example 7, the carrier matrix was loaded into the housing along with the densifying agent and sealed. Bolas with lecithin gels were loaded in slightly lower quantities due to their lower density and tendency to occupy more space. Those skilled in the art will understand that a larger surface area for diffusion can affect the mass diffusing out of the bolus. Here, an attempt was made to minimize the variation between boluses in terms of the surface area in contact with the TBM inside the housing by the loaded matrix. Release test

[0357] As described in Example 7, each bolus was placed in 1 L of 0.02 M phosphate buffer, and the amount of TBM released from the bolus daily was quantified. result

[0358] In boluses containing lecithin-based gels (Figure 4), the release rate was directly proportional to the amount of TBM loaded in the gel (Figure 5). However, the release profile was first-order for gels loaded with 60% or 70% TBM. A near-zero-order release profile was observed in gels loaded with 45% TBM. This behavior was observed over a long period of more than 4 months (Figure 6).

[0359] Similarly, PMMA gels (Figure 7) also exhibited TBM concentration-dependent release (Figure 8). Interestingly, we observed that a PMMA gel with a 45% TBM load released a minimum of TBM (<10 mg / day) over three months (Figure 9), suggesting that PMMA could be an excellent packing material for TBM-loaded boluses. Nevertheless, PMMA can be used as a stabilizer to modulate release from other matrices, such as stearic acid-based matrices, which provides limited control over release in the absence of the stabilizer, as will be discussed later. Example 11

[0360] Bolas with different melting point (MP) waxes loaded with TBM were also prepared. Microcrystalline wax (MCW) supplier Alchemy agency (specific gravity 0.92 at 20°C, melting point 80°C) with a melting point of approximately 80°C, stearic acid (purity 95%) with a melting point of approximately 70°C, and eicosane (purity >95%) with a relatively low melting point of approximately 40°C were used. In the case of the eicosane system, the aim was primarily to achieve temperature-dependent release. Bolus preparation

[0361] Different boluses were prepared using the wax / TBM matrices listed in Table 5. Briefly, in the MCW system, the wax was first melted and maintained at approximately 100°C, to which pre-melted TBM (kept at room temperature) was added. This allowed the temperature to decrease to approximately 70°C. Here, care was taken to pour the mixture into the housing body in a clear liquid state before any of them solidified, in particular, since multiple boluses were prepared from the same melt, in order to avoid non-uniformity of the matrix load. Those skilled in the art will understand that the pouring temperature or stirring / precipitation prior to pouring in such a system can affect their release profile by affecting their solid state. Stearic acid was melted and maintained at approximately 90°C, to which TBM was added. As discussed earlier for the MCW system, care was taken to pour it in a clear liquid state. Eicosane was melted and maintained at approximately 60°C, to which TBM was added. Here, the mixture was also above 40°C and was in a clear liquid state when poured into the housing. The bolus housing was the same as that described in Example 7. In all cases, the wax / TBM system was permitted to solidify before adding a densifying agent and sealing them with caps, as described in Example 7.

[0362] Table 5. Composition of support / matrix in each bolus. TBM = Tribromomethane; MCW = Microcrystalline wax. [Table 5] Release test

[0363] As described in Example 7, each bolus was placed in 1 L of 0.02 M phosphate buffer, and the amount of TBM released from the bolus daily was quantified. Specific to eicosane-based boluses, the boluses were kept at room temperature for approximately two weeks, and then moved to a 40°C incubator. The release of TBM from the boluses during this period, encompassing changes in the release pattern throughout the transition, was recorded. result

[0364] Both MCW systems exhibited release, but it was observed that a TBM loading of less than 70% provides more desirable control over TBM release (Figure 10). The MCW system with a 65% TBM loading showed consistent release of approximately 150 mg per day over 80 days (Figure 11). Furthermore, release can also be modulated based on the mass loading of MCW / TBM in 35% MCW w / w / TBM 65% w / w systems in the bolus, which was observed, and this is a desirable characteristic of such systems. Herein, boluses can be designed to deliver the desired dose to animals based on their weight by simply varying the loading of the matrix in the bolus.

[0365] In stearic acid systems, the release rate was considerably higher than that of MCW systems at the same loading, and release was in the range of approximately 500 mg per day (Figure 13). In some applications, a higher release rate may be desirable. However, at 65% TBM loading, the stearic acid system did not provide as much control over release as the MCW system (Figures 10 and 13).

[0366] For eicosane systems (Figure 14), it was observed that no significant TBM release occurs at room temperature, however, significant release occurs when the bolus is heated to 40°C. Without being bound by theory, it is believed that melting of the carrier accounts for the temperature-dependent release. Those skilled in the art will appreciate that such systems can be advantageous because they do not release a significant amount of active substance during storage, and only release a significant amount of active substance once administered to an animal. Furthermore, release from such systems can be modulated by including excipients such as EC, lecithin, MCW, or SAIB that can further control the release of active agents such as TBM.

[0367] It is also interesting to note that the release of TBM from wax systems with the same TBM load increased as the melting point of the support wax decreased. At a 65% TBM load into the wax system, MCW (MP = approximately 80°C) provided better control over release compared to stearic acid (MP = approximately 70°C), which in turn provided better control over release from eicosane (MP = approximately 80°C) as the support. Example 12

[0368] Using MCW as the carrier, boluses without a housing were also prepared. Bolus preparation

[0369] The bolus housing (when used) was the same as that described in Example 7. Here, MCW matrices loaded with 65% or 45% TBM loads were prepared and loaded either into boluses or boluses without housings (Table 6). Briefly, boluses with housings were prepared as described in Example 11. Boluses without housings were prepared using 3D printed molds. Alternatively, once the matrix has solidified, the housing can be cut off and removed. Since the boluses were dense enough to sink in water, densifying agents were not included in these tests. However, if required in equivalent boluses for administration to animals, those skilled in the art could easily include further densifying agents.

[0370] Table 6. Composition of support / matrix in each bolus. TBM = Tribromomethane; MCW = Microcrystalline wax [Table 6] Release test

[0371] As described in Example 7, each bolus was placed in 1 L of 0.02 M phosphate buffer, and the amount of TBM released from the bolus daily was quantified. Potentially, due to the high density of TBM, which formed a more significant proportion of the bolus in the absence of a housing, all housing-free boluses also sank into the dissolution medium. result

[0372] The boluses without housings appeared to release TBM at approximately 2–3 times the rate of the equivalent boluses with housings (Figures 15 and 16). Nevertheless, the boluses without housings were stable in the dissolution medium, did not break during testing, and suggested that they possess satisfactory mechanical properties, at least when MCW is used. Based on these results, at least other waxes with higher melting points than MCW may also be used in boluses without housings. Bolus without housings may offer advantages in using reduced materials for potentially simpler production methods. Bolus without housings may also be advantageous when high release rates are desired. Housings may be advantageous when greater modulation of the release rate is required. Example 13

[0373] The inventors observed that different excipients / carriers provide different levels of control over TBM release. Specifically, EC, lecithin, SAIB, and MCW were observed to provide significant control over TBM release; a desirable characteristic in sustained-release formulations. Therefore, EC, lecithin, SAIB, and MCW act as stabilizers for TBM release. Here, lecithin was used as an example of a stabilizer for a system that improves control over TBM release compared to stearic acid systems. Bolus preparation

[0374] Matrices with 65% TBM loaded with stearic acid were prepared with or without 5% lecithin (Table 7). Lecithin inclusion was compensated for by a decrease in stearic acid concentration. The bolus housing was the same as that described in Example 7. Here, the bolus was assembled as described in Example 11. In the case of the matrix with lecithin, the lecithin was first dissolved in TBM, and then the solution was added to the molten stearic acid.

[0375] Table 7. Composition of carrier / matrix in each bolus. TBM = Tribromomethane. [Table 7] Release test

[0376] As described in Example 7, each bolus was placed in 1 L of 0.02 M phosphate buffer, and the amount of TBM released from the bolus daily was quantified. result

[0377] It was observed that the inclusion of only 5% lecithin could stabilize the release profile of the stearic acid system loaded with 65% TBM (Figure 17). This suggests that similar control may be achieved among other stabilizer excipients, such as SAIB, EC, lecithin, and MCW. Example 14

[0378] A bolus may contain more than one active ingredient. The inclusion of more than one active agent offers advantages, at least in terms of reduced application of the active agent. Furthermore, they may potentially offer advantages from a formulation standpoint. Here, boluses were prepared containing the additional active agents monensin (sodium salt, purity ≥90%), phloroglucinol (dihydrate, purity ≥98%), ketoprofen (purity ≥98%), and albendazole (purity ≥98%). Monensin is known to exhibit anti-methane activity (Cooke RF et al. (2024) Transl Anim Sci, Mar 9;8:txae032). Phloroglucinol is a hydrogen sequestering agent that may potentially help divert excess hydrogen to acetate in rumens where anti-methanogenic agents such as haloform have suppressed methane production (which uses hydrogen), and is also known to reduce methane production (Sarwono KA et al. (2019) Tropical Animal Science Journal, 42(2):121-127). Ketoprofen is a common anti-inflammatory agent used in cattle. Albendazole is a common anthelmintic used in cattle. Bolus preparation

[0379] EC / HPMC / TBM matrices loaded with 60% TBM, with or without inclusion of 2% or 5% monensin, phloroglucinol, albendazole, and ketoconazole, were prepared as detailed in Table 8. Inclusion of further active agents in the formulations was compensated for by a reduction in HPMC. Since EC is established to provide stability to the release profile, the ratio of EC to TBM was kept constant here. These active agents were first added to the TBM. The matrices and housings were prepared as described in Example 7.

[0380] Table 8. Composition of support / matrix in each bolus. TBM = Tribromomethane; EC = Ethylcellulose, HPMC = Hydroxypropylmethylcellulose. [Table 8] Release test

[0381] As described in Example 7, each bolus was placed in 1 L of 0.02 M phosphate buffer, and the amount of TBM released from the bolus daily was quantified. result

[0382] The inclusion of additional active agents produced only minor changes in the release profile (Figures 18–21). Example 15

[0383] The effect of casing crystallinity on the release rate was evaluated. Bolus preparation

[0384] The bolus housing was the same as that described in Example 7, with the exception that a bolus housing with a high crystalline PLA content and a high amorphous crystalline content was prepared. Those skilled in the art will know techniques for influencing the crystalline polymer content, such as annealing. Here, an MCW matrix loaded with 65% TBM up to a total mass of 60 g was prepared as described in Example 11. Release test

[0385] As described in Example 7, each bolus was placed in 1 L of 0.02 M phosphate buffer (pH=6.5), and the amount of TBM released from the bolus daily was quantified. result

[0386] Bolus with a high crystalline PLA content exhibited a significantly higher release rate than boluses with a low crystalline PLA content. This suggests that the crystallinity of the casing affects the release rate. Claims of the present invention

[0387] 1. A bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: Core containing a methane inhibitor; and Housing surrounding the core, Here, the housing is configured such that its permeability to the methane inhibitor increases when the housing is exposed to the lumen of a living animal; and / or

[0388] Here, at least a portion of the housing is configured to form one or more openings when the housing is exposed to the lumen of a living animal, allowing for increased release of methane inhibitor from within the bolus through the opening(s).

[0389] 2. A bolus in accordance with Claim 1, wherein the housing comprises one or more openings, the openings being filled with and / or covered with a material that melts, dissolves or collapses in the rumen of a living animal.

[0390] 3. A bolus in accordance with Claim 1, wherein a portion of the housing, preferably an opening(s) formed by a portion of the housing, is made of a material that melts, dissolves, or collapses in the rumen of a living animal.

[0391] 4. A bolus of claim 2 or 3, wherein the material to be melted is a compound selected from the group consisting of hydrogels, oleogels, organogels, phase change materials (PCMs), fatty acids, alkanes, alkenes, gelling agents, waxes, L-alanine amino acids, L-alanine amino acid derivatives, poly(methyl methacrylate) (PMMA), (1,3:2,4) dibenzylidene sorbitol (DBS), hydroxystearic acid, paraffin wax, gelatin, 1-tetradecanol, polyethylene glycol, octadecane, nonadecane, eicosane, Pluronic® polymer or mixtures of Pluronic®, emulsifiers, sucrose acetate isobutyrate (SAIB), derivatives of the above, and one or more combinations of the above compounds, wherein the compound or combination of compounds has a melting temperature between 28°C and 42°C, more preferably between 28°C and 35°C.

[0392] 5. A bolus of claim 2 or 3, wherein the material to be dissolved is preferably a water-soluble material selected from the group consisting of polymers, polyols, sugars, polyamides, salts, and cellulose acetate.

[0393] 6. A bolus of claim 2 or 3, wherein the material to be disintegrated is a compound selected from the group consisting of cellulose, polyhydroxyalkanoate (PHA), poly(butylene succinate-co-adipate) (PBSA), and a mixture of two or more of the above.

[0394] 7. A bolus conforming to any one of claims 1-6, wherein the largest opening formed on the housing when the housing is exposed to the lumen of a living animal has a maximum diameter of 2 mm.

[0395] 8. A bolus according to any one of claims 1-7, wherein the methane inhibitor is selected from haloform.

[0396] 9. A bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: The core contains a methane inhibitor in microencapsulated particles, and the microencapsulated particles are dispersed in a composition that includes a carrier and optionally a dispersant. and a housing that covers at least a portion of the core.

[0397] 10. A bolus in accordance with claim 9, wherein microencapsulated particles can be produced by microencapsulation of the particles in at least one encapsulating agent, preferably wherein the encapsulating agent is selected from the group consisting of polymers, surfactants, emulsifiers, gelatin-sorbitol mixtures, gelatin-starch syrup, and mixtures thereof, more preferably wherein the encapsulating agent is selected from the group consisting of PLA, PBSA, PBS, PHBV, PVA, PBAT, PCL, PDLA, gelatin-sorbitol mixtures, gelatin-starch syrup, and mixtures containing two or more of the compounds described above.

[0398] 11. A bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: A carrier and microencapsulated particles, where the microencapsulated particles contain a methane inhibitor and are dispersed within the carrier. Preferably, the carrier has a porous structure such as mesoporous silica.

[0399] 12. A bolus according to either claim 9 or 11, wherein the microencapsulated particles are microencapsulated using compounds selected from the group consisting of hydrophilic materials, gelatin, zein, methylcellulose and poly(N-isopropylacrylamide) (PNIPAM) microgels, starch, cyclodextrin and two or more combinations of the compounds described above.

[0400] 13. A bolus according to any one of claims 9 to 12, wherein the carrier comprises a compound selected from the group consisting of silica, cellulose and activated carbon, gelatin, chitosan, poly(lactic acid-coglycolic acid) (PLGA), cyclodextrin, collagen, polyalpha-hydroxyester, hydroxyalkanoate and dioxane, starch, gluten, zein, polyethylene, polypropylene, polyamide, polyethylene terephthalate and ethylene vinyl acetate.

[0401] 14. A bolus according to any one of claims 9 to 13, wherein the microencapsulated particles have an average diameter of 50 nm to 2 mm, preferably an average diameter of 1 μm to 1000 μm.

[0402] 15. A bolus in accordance with any one of claims 9-14, wherein the bolus is configured to release a methane inhibitor over a period of at least three months.

[0403] 16. A bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal. Here, the bolus includes a core, and the core includes a methane inhibitor and a carrier. The bolus does not include the housing.

[0404] 17. A bolus in accordance with Claim 16, wherein the core comprises a compound selected from the group consisting of PLA, PBSA, PBS, PHBV, PVA, PBAT, PCL, PDLA, epoxy chain extenders, magnesium silicate, cellulose materials, ethylcellulose, hydroxypropyl methylcellulose (HPMC), fumed silica, gelatin, wax, caster wax, paraffin wax, silica, and combinations thereof.

[0405] 18. A bolus in accordance with either claim 16 or 17, wherein the bolus has a Shore D hardness of at least 20.

[0406] 19. A bolus in accordance with any one of claims 16-18, wherein the bolus is in the form of a pellet, pill, lozenge, or tablet.

[0407] 20. A bolus conforming to any one of claims 16-19, wherein the bolus is configured to dissolve in the rumen of a ruminant over a period of less than 48 hours.

[0408] 21. A bolus according to any of the preceding claims, wherein the bolus preferably comprises a chelating agent selected from the group of chelating agents consisting of fumaric acid, sodium fumarate, phenol compounds, phloroglucinol, gallic acid, resorcinol, catechol, hydroquinone, and pyrogallol.

[0409] 22. A bolus according to any of the preceding claims, wherein the methane inhibitor is selected from the group consisting of bromoform, monensin, lecithin, lauric acid, myristic acid, linoleic acid, and phospholipids, preferably wherein the phospholipid comprises one or more polyunsaturated fatty acids, more preferably wherein the phospholipid comprises one or more fatty acids selected from 18-carbon polyunsaturated fatty acids (C18 PUFAs), palmitic acid, stearic acid, and oleic acid.

[0410] 23. A bolus for administration to a ruminant, wherein the bolus is configured to release a methane inhibitor in the animal, wherein the bolus comprises: Cores comprising methane inhibitors and carriers; and Housing including the aforementioned core: Here, the methane inhibitor is selected from the group consisting of monensin, lauric acid, myristic acid, and linoleic acid; the housing includes at least one opening that exposes the core to the environment surrounding the bolus.

[0411] 24. A bolus according to any one of the preceding claims, wherein the bolus comprises a housing, wherein the housing comprises a stabilizer, preferably selected from the group consisting of surfactants, plasticizers, phthalates, and triglycerides, more preferably, wherein the stabilizer is selected from the group consisting of lecithin, nitriles, and triacetin.

[0412] 25. A bolus conforming to any one of the preceding claims, wherein the core of the bolus comprises at least one filler, Preferably, the filler is a stabilizer, and preferably, the filler is selected from the group consisting of gelatin, milk, milk derivatives, infant formula, powdered milk, triglycerides, medium-chain triglycerides and their oils, ethanol, lecithin, tween, xanthan gum, cellulose derivatives, alkylcellulose, ethylcellulose, hydroxypropyl methylcellulose (HPMC), zein, and surfactants.

[0413] 26. A bolus as defined in any one of the preceding claims, wherein the bolus comprises an active agent in addition to or instead of the methane inhibitor, wherein the active agent is selected from the group consisting of anti-inflammatory agents, analgesics, antibiotics, and anthelmintics; more preferably, wherein the active agent is selected from the group consisting of meloxicam, ketoprofen, penicillin, tetracycline, macrolides, monensin, ceftiofur, florfenicol, tilmicosin, enrofloxacin, tulathromycin, and albendazole.

[0414] 27. A bolus in accordance with any one of claims 1 to 26 for use in the treatment of ruminants.

[0415] 28. A bolus in accordance with any one of claims 1 to 26 for use in reducing methane emissions in ruminants.

[0416] 29. A method for treating an animal, comprising administering to the animal a bolus as defined in any one of the preceding claims, wherein the animal is preferably a ruminant such as a cattle.

[0417] 30. How to manufacture a bolus that follows any one of claims 1-28, including the following steps: (a) Provide housing; (b) Multiple openings are created in the wall of the housing, where each opening has a maximum diameter of 3 mm; (c) sealing the opening of the housing with the material which melts, dissolves, and / or disintegrates in the rumen of a living animal; and (d) Filling the housing with the inhibitor.

[0418] 31. A methane inhibitor for use in reducing methane emissions from ruminants, wherein the methane inhibitor is selected from the group consisting of bromoform, monensin, lecithin, lauric acid, myristic acid, linoleic acid, and phospholipids, preferably wherein the phospholipid comprises one or more polyunsaturated fatty acids, more preferably wherein the phospholipid comprises one or more fatty acids selected from 18-carbon polyunsaturated fatty acids (C18 PUFAs), palmitic acid, stearic acid, and oleic acid.

[0419] 32. A method for treating ruminants to reduce methane emissions from them, comprising administering to the animals a methane inhibitor selected from the group consisting of bromoform, monensin, lecithin, lauric acid, myristic acid, linoleic acid, and phospholipids, preferably wherein the phospholipid comprises one or more polyunsaturated fatty acids, and more preferably wherein the phospholipid comprises one or more fatty acids selected from 18-carbon polyunsaturated fatty acids (C18 PUFAs), palmitic acid, stearic acid, and oleic acid.

Claims

1. A bolus configured to release a methane inhibitor in a ruminant, wherein the bolus comprises: core; A housing that covers at least a portion of the core; Here, the core comprises at least one methane inhibitor and at least one further active agent.

2. The bolus according to claim 1, wherein the further active agent is selected from the group consisting of methane inhibitors, hydrogen sequestering agents, anti-inflammatory agents, analgesics, anthelmintics, nonsteroidal anti-inflammatory drugs (NSAIDs), antibiotics, growth promoters, milk secretion promoters, sustained release enhancers, antibacterial agents, ketosis inhibitors, and combinations thereof.

3. The bolus according to claim 1 or 2, wherein the further active agent is selected from the group consisting of monensin, phloroglucinol, albendazole, ketoconazole, lecithin, and combinations thereof.

4. The bolus according to any one of claims 1 to 3, wherein the further active agent is monensin.

5. A bolus configured to release a methane inhibitor in a ruminant, wherein the bolus comprises: A core containing a methane inhibitor; and a housing covering at least a portion of the core. Here, the housing is configured such that, when exposed to a phosphate buffer (pH: 6.5, 0.02 M) at 40°C without stirring, its permeability to the methane inhibitor increases by at least 50% relative to its permeability on the same day after exposure to the phosphate buffer (pH: 6.5, 0.02 M) at the reference temperature without stirring for the same duration. Here, permeability was evaluated based on the release rate of the methane inhibitor on day 14; Here, the reference temperature is 20°C.

6. The bolus according to claim 5, wherein the reference temperature is 25°C.

7. A bolus configured to release a methane inhibitor in a ruminant, wherein the bolus comprises: A core comprising a methane inhibitor dispersed in or forming part of one or more hydrogels, oleogels, or organogels; and A housing that covers at least a portion of the core.

8. The bolus according to claim 7, wherein the core comprises a phospholipid and / or acrylate polymer.

9. The bolus according to claim 7 or 8, wherein the core comprises lecithin and / or poly(methyl methacrylate) (PMMA).

10. A bolus according to any one of claims 1 to 9, wherein the housing surrounds the core.

11. A bolus according to any one of claims 1 to 10, wherein the housing comprises one or more polymers selected from the list consisting of high-density polyethylene (HDPE), polypropylene (PP), polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), poly-D,L-lactic acid (PDLLA), polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), talc-filled poly(D-lactide) (TALC PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), combinations thereof, and copolymers thereof.

12. The bolus according to any one of claims 1 to 11, wherein the housing comprises one or more biodegradable polymers.

13. The bolus according to any one of claims 1 to 12, wherein the housing comprises one or more polymers selected from the list consisting of polylactic acid, polybutylene adipate terephthalate, combinations thereof, and copolymers thereof.

14. A bolus according to any one of claims 1 to 13, wherein the housing completely encloses the core.

15. A bolus according to any one of claims 1 to 14, wherein the housing is 5-100% or 10-100% PLA w / w.

16. A bolus according to any one of claims 1 to 15, wherein the housing is 20-100% or 30-90% PBAT w / w.

17. The bolus according to any one of claims 1 to 16, wherein the housing has a material thickness less than about 2 mm, preferably in the range of about 0.3 to 1.8 mm, and more preferably in the range of about 0.3 to 1.5 mm.

18. A bolus according to any one of claims 1 to 17, wherein the core and housing have a weight ratio of approximately 3 to approximately 6:1, approximately 4 to approximately 5:1, or approximately 4.6:

1.

19. A bolus according to any one of claims 1 to 18, wherein the bolus has a Shore D hardness of at least 20.

20. A bolus configured to release a methane inhibitor in a ruminant, wherein the bolus comprises: The core, where the core contains a methane inhibitor and a carrier, and the bolus does not contain a housing.

21. The core consists of polycaprolactone (PCL), polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), polylactic acid (PLA), polylactic acid, poly-d-lactic acid, poly-L-lactic acid, poly-D,L-lactic acid (PDLLA), polylactide-co-glycolide, lignin, polybutylene adipate terephthalate (PBAT), styrene-acrylic copolymer (e.g., Joncryl®), and talc-filled poly(D-lactide) (TALC). PDLA), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), polyvinyl alcohol (PVA), epoxy chain extender, magnesium silicate, cellulose material, ethylcellulose, hydroxypropyl methylcellulose (HPMC), fumed silica, gelatin, wax, castor wax, paraffin wax, silica, hydrophilic silica, microcrystalline wax, methylcellulose, starch, polyethylene glycol, polyvinyl alcohol, hydroxyethylcellulose, carboxymethylcellulose, Solplus, poly(acrylic acid), poly(vinylpyrrolidone), poly(vinyl alcohol), poly(acrylamide), poly(2-hydroxypropyl methacrylamide), poly(N,N-dimethylacrylamide), poly([2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide), poly(2-(methacryloyloxy)ethyl phosphorylcholine), poly(carboxybetaine methacrylamide), poly(ethylene glycol), poly(ethyleneimine), poly(sarcosine), poly(2-methyl-2-oxazoline), polyaminoesters, polyesteramides, polyphosphoesters, poly(l-lysine), poly(l-proline), polyphosphazene, dextran, sodium alginate, gelatin, agarose, carrageenan, guerlain, xanthan gum, urea, sucrose, beeswax, polyethylene glycol (PEG), starch A bolus according to any one of claims 1 to 20, comprising one or more materials selected from the group consisting of sodium benzoate, sodium croscarmellose, crospovidone, carboxymethylcellulose (CMC), hydroxyethylcellulose (HEC), carrageenan, guar gum, xanthan gum, sodium alginate, locust bean gum, polyvinylpyrrolidone (PVP), polyvinylpyrrolidone-vinyl acetate copolymer (PVP / VA), polyacrylic acid and / or copolymer variants thereof, polyisobutylene, vinyl ethyl acetate (EVA), functional waxes having a melting point less than about 120°C, their derivatives, combinations thereof, and their copolymers.

22. The bolus according to any one of claims 1 to 21, wherein the core comprises one or more materials selected from the group consisting of ethylcellulose, hydroxypropyl methylcellulose (HPMC), combinations thereof, and copolymers thereof.

23. A bolus according to any one of claims 1 to 22, wherein the core comprises microcrystalline wax.

24. The bolus according to any one of claims 1 to 23, wherein the methane inhibitor is a haloform, preferably a bromoform.

25. The bolus according to claim 24, wherein a haloform, preferably bromoform, is contained in the core of the bolus of the present disclosure in an amount between 10 wt% and 80 wt%, preferably between 15 wt% and 70 wt%.

26. The bolus according to claim 25, wherein bromoform is not a bromoform-rich seaweed extract.

27. The bolus according to any one of claims 1 to 26, wherein the bolus further comprises a high-densification agent.

28. A bolus according to any one of claims 1 to 27, wherein the bolus contains the methane inhibitor bromoform and is adapted to reach a maximum release rate of approximately 0.1 to approximately 0.5 g per day, more preferably approximately 0.2 g per day.

29. A bolus according to any one of claims 1 to 28, wherein the bolus comprises a therapeutically effective amount of a methane inhibitor and the following: • Approximately 5% to 15% (w / w) housing; • Approximately 20 to 55 (w / w) cores; and • High-densification agent with a density of approximately 30 to 75 (w / w).

30. A method for administering a methane inhibitor to a ruminant, comprising administering a bolus according to any one of claims 1 to 29 to the rumen of the ruminant.

31. A method for reducing methane production in the rumen of a ruminant, comprising administering a bolus according to any one of claims 1 to 29 to the rumen of the ruminant.

32. The method of creating a bolus includes the following: Selecting the core and housing, Inserting the core into the housing, Optionally, sealing the housing to enclose or substantially enclose the core inside the housing; Here, the core contains a methane inhibitor.

33. The method according to claim 32, wherein a molten core is poured into a housing and solidifies within the housing.

34. The method according to claim 33, wherein the housing surrounds the core after the core has solidified.

35. A packaged bolus according to any one of claims 1 to 29, or a prepared bolus according to claims 32 and 33, wherein the bolus is packaged in a material containing an acrylate polymer.

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