阅读说明：本技术 非洲猪瘟病毒和经典猪瘟病毒的化学缓疫 (Chemical pestilence of African swine fever virus and classical swine fever virus ) 是由 梅甘·C·尼德沃德 雷蒙德·R·R·罗兰 卡桑德拉·琼斯 史蒂文·S·德里茨 杰森·C·伍德 于 2019-03-01 设计创作，主要内容包括：提供了抑制在动物饲料、饲料成分和宠物食品中的非洲猪瘟病毒和/或经典猪瘟病毒的传播的方法。方法使用总体上安全的化学缓疫剂,诸如中链脂肪酸。化学缓疫剂当以比抑制其它微生物的现有方法低得多的包含率引入到饲料或饲料成分中时是有效的。方法特别适用于将运输和储存数天或数周的动物饲料、饲料成分或宠物食品的后加工处理。(Methods of inhibiting the transmission of African swine fever virus and/or classical swine fever virus in animal feed, feed ingredients, and pet food are provided. The method uses a generally safe chemical buffer, such as a medium chain fatty acid. Chemical buffers are effective when introduced into feed or feed ingredients at a much lower inclusion rate than existing methods of inhibiting other microorganisms. The method is particularly suitable for post-processing treatment of animal feed, feed ingredients or pet food which is to be transported and stored for days or weeks.)

1. A method of inhibiting african swine fever virus and/or classical swine fever virus in an animal feed or animal feed ingredient, the method comprising:

introducing a chemical buffer to the feed or feed ingredient, the chemical buffer comprising a medium chain fatty acid and/or an essential oil,

wherein the chemical retardant is introduced at an inclusion rate of about 0.125 wt% to less than 2 wt%, based on the total weight of the animal feed or feed ingredient taken as 100 wt%.

2. The method of claim 1, wherein the medium chain fatty acid is selected from the group consisting of: caproic acid, caprylic acid, capric acid, lauric acid and mixtures thereof.

3. The method of claim 1 or 2, wherein the chemical retardant is a blend of two or more medium chain fatty acids.

4. The method of any one of claims 1-3, wherein the chemical retardant comprises a blend of medium chain fatty acids including hexanoic acid, octanoic acid, and decanoic acid.

5. The method of any one of claims 1-4, wherein the chemical retardant comprises a blend of medium chain fatty acids comprising about equal parts of hexanoic acid, octanoic acid, and decanoic acid.

6. The method according to any one of claims 1 to 5, wherein the essential oil is selected from the group consisting of: garlic oleoresin, turmeric oleoresin, capsicum oleoresin, rosemary extract, wild oregano essential oil and mixtures thereof.

7. The method of any one of claims 1 to 6, wherein the chemical retardant comprises a blend of medium chain fatty acids and essential oils.

8. The method of any one of claims 1 to 7, wherein the animal feed or animal feed ingredient is selected from the group consisting of: complete pig feed, blood meal, pork bone meal (MBM), spray-dried animal plasma, feather meal, poultry blood meal, poultry by-product meal, vitamin D, lysine hydrochloride, choline chloride and soybean meal.

9. The method of any one of claims 1 to 7, wherein the animal feed or animal feed ingredient is dry pet kibble, the method further comprising applying sodium bisulfate on the surface of the dry pet kibble.

10. The method of claim 9, wherein the sodium bisulfate is applied to the surface of the dry pet treat at a inclusion rate of from about 0.1% to about 2% by weight, based on the total weight of the animal feed or feed ingredient taken as 100% by weight.

11. Chemical buffer for use in inhibiting African swine fever virus and/or classical swine fever virus in an animal feed or animal feed ingredient, the chemical buffer comprising medium chain fatty acids and/or essential oils.

12. The chemical retardant of claim 11, wherein the medium chain fatty acid is selected from the group consisting of: caproic acid, caprylic acid, capric acid, lauric acid and mixtures thereof.

13. A chemical buffering agent according to claim 11 or 12, wherein the chemical buffering agent is a blend of two or more medium chain fatty acids.

14. A chemical buffering agent according to any of claims 11 to 13, wherein the chemical buffering agent comprises a blend of medium chain fatty acids including hexanoic acid, octanoic acid and decanoic acid.

15. A chemical buffering agent according to any one of claims 11 to 14, wherein the chemical buffering agent comprises a blend of medium chain fatty acids comprising about equal parts of hexanoic, octanoic and decanoic acids.

16. A chemical retardant according to any one of claims 11 to 15 wherein the essential oil is selected from the group consisting of: garlic oleoresin, turmeric oleoresin, capsicum oleoresin, rosemary extract, wild oregano essential oil and mixtures thereof.

17. A chemical buffer as claimed in any of claims 11 to 16 wherein the chemical buffer comprises a blend of medium chain fatty acids and essential oils.

18. A chemical deterrent according to any of claims 11-17, wherein the chemical deterrent comprises a blend of essential oils comprising about equal parts of garlic oleoresin, turmeric oleoresin, capsicum oleoresin, rosemary extract, and wild oregano essential oil.

19. A treated animal feed or animal feed ingredient resistant to african swine fever virus and/or classical swine fever virus comprising from about 0.125 wt% to less than 2 wt% of a chemical buffering agent comprising medium chain fatty acids and/or essential oils, based on the total weight of the animal feed or feed ingredient taken as 100 wt%, wherein the animal feed or animal feed ingredient is selected from the group consisting of: complete pig feed, blood meal, pork bone meal (MBM), spray-dried animal plasma, feather meal, poultry blood meal, poultry by-product meal, vitamin D, lysine hydrochloride, choline chloride, soybean meal, dry pet roughage, and mixtures thereof.

20. The treated animal feed or animal feed ingredient resistant to African swine fever virus and/or classical swine fever virus according to claim 19, wherein the animal feed or animal feed ingredient is a dry pet treat further comprising sodium bisulfate on the surface of the dry pet treat.

Technical Field

The present invention relates broadly to methods of inhibiting african swine fever virus and/or classical swine fever virus in animal feed, feed ingredients and pet food.

Background

Disclosure of Invention

The present application describes methods for inhibiting african swine fever virus and/or classical swine fever virus in an animal feed or animal feed ingredient. The method includes introducing a chemical buffering agent into the feed or feed ingredient. Chemical buffers include (consist essentially of or even consist of): medium chain fatty acids and/or essential oils, and a chemical buffering agent is introduced into the feed or feed ingredient at a inclusion rate of less than 2 wt% (but typically at least about 0.125 wt%), based on the total weight of the animal feed or feed ingredient taken as 100 wt%.

In another embodiment, there is provided a chemical buffering agent for use in inhibiting african and/or classical swine fever virus in an animal feed or animal feed ingredient. The chemical buffer comprises medium chain fatty acids and/or essential oils.

Also described herein are treated animal feed and/or animal feed ingredients that are resistant to African swine fever virus and/or classical swine fever virus. The feed or feed ingredient comprises from about 0.125 wt% to less than 2 wt% of the chemical buffering agent, based on the total weight of the animal feed or feed ingredient taken as 100 wt%. The chemical buffering agent comprises medium chain fatty acid and/or essential oil. Exemplary treated animal feed or animal feed ingredients for use in the present invention include complete pig feed, blood meal, pork bone meal (MBM), spray dried animal plasma, feather meal, poultry blood meal, poultry by-product meal, vitamin D, lysine hydrochloride, choline chloride, soy meal, dry pet roughage, and mixtures thereof.

Drawings

This patent or application document contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the office upon request and payment of the necessary fee.

Figure (fig.)1 is a graph showing a dose-response inactivation curve for ASFV (strain BA71V) exposed to different concentrations of a 1:1:1MCFA blend, with data shown as titers after exposure to MCFA concentrations of 0.125% to 2.0%, and the percentage of reduction in virus concentration compared to positive controls;

figure 2 is a series of graphs showing detection of the ASFV Georgia 2007 genome over the course of a 30-day cross-border model, with data shown as mean cycle threshold (Ct) values at 1, 8, 17, and 30 days post-inoculation in duplicate; and

FIG. 3 is a graph showing the amount of ASFV DNA in an untreated control (open bar) and a sample treated with MCFA at 28dpi (black bar) measured by qPCR at the end of a 30-day cross-border model;

FIG. 4 is a graph showing dose-response inactivation curves for CSFV (Brescia isolate) exposed to different concentrations of a 1:1:1MCFA C6: C8: C10 blend, where the data are shown as titers after exposure to MCFA concentrations of 0.125% to 2.0%, and as a percentage of reduction in virus concentration compared to a positive control; and

FIG. 5A is a positive control image from an experiment of indirect fluorescent antibody detection of CSFV Brescia on porcine kidney cells exposed or not exposed to MCFA;

FIG. 5B is an image of an experiment from indirect fluorescent antibody detection of CSFV Brescia on porcine kidney cells treated with 0.625% MCFA; and

figure 5C is a negative control image of an experiment derived from indirect fluorescent antibody detection of CSFV Brescia on porcine kidney cells with or without exposure to MCFA.

Detailed description of the preferred embodiments

The present invention relates generally to methods of inhibiting African Swine Fever Virus (ASFV) and/or Classical Swine Fever Virus (CSFV) in animal feed, feed ingredients and pet food. More particularly, the invention relates to the use of chemical buffering agents for the inhibition of ASFV and/or CSFV in various types of animal and pet food ingredients, as well as in complete feed diets and pet food products. Generally, the chemical buffering agent includes a medium chain fatty acid and/or an essential oil. As used herein, "inhibit" or "inhibition" refers to a decrease in the measurable level of a microorganism of interest (i.e., ASFV or CSFV) or a decrease in the growth rate of the microorganism as compared to an untreated control. In one or more embodiments, the methods according to the invention employ an effective amount of a chemical buffering agent to inhibit ASFV and/or CSFV in an animal feed or animal feed ingredient, e.g., to a concentration below the level detected by RT-PCR and/or virus isolation in cell culture. As used herein, "effective amount" refers to an amount of active compound (e.g., medium chain fatty acids and/or essential oils) that is capable of providing a bioavailable level sufficient to achieve the desired performance improvement. In a preferred embodiment, the method according to the invention is advantageously suitable for the transport and storage of animal feed ingredients.

Chemical buffering agents for use in one or more embodiments of the present invention may include medium chain fatty acids and/or essential oils. In one or more embodiments, the chemical buffering agent comprises (consists essentially of or even consists of): medium chain fatty acids, and more specifically at least one medium chain fatty acid. Medium chain fatty acids are acids with an aliphatic tail of 6 to 12 carbon atoms. In one or more embodiments, medium chain fatty acids useful herein include caproic acid, caprylic acid, capric acid, and/or lauric acid. Thus, in certain embodiments, the chemical buffering agent is selected from the group consisting of: caproic acid, caprylic acid, capric acid, lauric acid and mixtures thereof. However, in certain other embodiments, the chemical buffering agent does not contain lauric acid. Thus, in such embodiments, the chemical buffering agent may be selected from the group consisting of: caproic acid, caprylic acid, capric acid and mixtures thereof. In one or more embodiments, a blend of medium chain fatty acids may be used. For example, in one or more embodiments, a blend of two or more medium chain fatty acids can be introduced into a feed or feed ingredient. In one or more embodiments, the blend of hexanoic acid, octanoic acid, and decanoic acid is introduced into the feed or feed ingredient in a weight ratio of about 1:1:1 (equal parts). This combination increases the solubility of medium chain fatty acids and is effective in inactivating virus and improving pig growth when administered orally. However, it is within the scope of the present invention to use blends comprising other weight ratios of hexanoic acid, octanoic acid and decanoic acid.

In one or more embodiments, the chemical buffering agent comprises (consists essentially of or even consists of): an essential oil, and more specifically at least one essential oil. Essential oils are concentrated hydrophobic liquids containing volatile aromatic compounds derived from plants. There are a variety of different essential oils that can be used in one or more embodiments of the present invention. A non-exclusive list of these essential oils includes: jojoba oil, Ajwain oil, angelica root oil, anise oil, asafetida, peruvian balsam, basil oil, bay oil, bergamot oil, black pepper, brussel oil, birch, camphor tree, hemp oil, caraway oil, cardamom oil, carrot seed oil, cedar oil, chamomile oil, calamus root, cinnamon oil, labdanum species, citron, citronella oil, sage, clove oil, coffee, coriander, chrysanthemum oil (chrysanthemum leaf oil), woody root, cranberry seed oil, piper cubeba, fennel/black seed oil, cypress, nutgrass, curry leaf, indene oil, dill oil, elecampane, eucalyptus oil, fennel seed oil, fenugreek oil, fir wood, shea butter, galangal, galbanum, geranium oil, maple oil, eucheuma, grapefruit oil, henna oil, ash oil, horseradish oil, jasmine oil, chamomile oil, sage oil, star anise oil, myrtle oil, geranium oil, maple oil, Bay (Laurus nobilis), lavender oil, ledum, lemon oil, lemon grass, lime, Litsea cubeba (Litsea cubeba) oil, eaglewood (Linaloe), citrus, marjoram, cajeput Tea tree oil (melaleuca Tea tree oil), bee balm oil (honey bee leaves), Mentha arvensis oil/peppermint oil, moringa oil, Savory (Mountain Savory), artemisia oil, mustard oil (essential oil), myrrh oil, myrtle, neem oil or neem oil, orange flower oil, nutmeg, orange oil, oregano oil, orris oil, sandalwood, parsley oil, patchouli oil, perilla essential oil, peppermint oil, pine leaf, juniper, roman, rose oil, rosemary oil, sage oil, peppermint oil, rose oil, sage oil, peppermint, Spikenard, spruce, star anise oil, citrus, tarragon leaf oil, tea tree oil, thyme oil, hemlock, turmeric, valerian, vetiver oil (vetiver oil), cedar, wintergreen, yarrow oil, ylang-ylang and zedoary. In one or more embodiments, the chemical buffering agent is selected from the group consisting of: garlic oleoresin, turmeric oleoresin, capsicum oleoresin, rosemary extract, wild oregano essential oil and mixtures thereof. In one or more embodiments, blends of essential oils may be used. For example, in one or more embodiments, a blend of two or more essential oils can be introduced into a feed or feed ingredient. In one or more embodiments, a blend of essential oils comprising equal parts of garlic oleoresin, turmeric oleoresin, capsicum oleoresin, rosemary extract and wild oregano essential oil is introduced into the feed or feed ingredient. However, it is within the scope of the present invention that blends comprising other weight ratios of garlic oleoresin, turmeric oleoresin, capsicum oleoresin, rosemary extract and wild oregano essential oil may be used.

In one or more embodiments, the following may be included (consisting essentially of or even consisting of): medium chain fatty acids and essential oils, and more specifically blends of one or more medium chain fatty acids and one or more essential oils, are used as chemical retardants. However, in certain other embodiments, no essential oils or other types of buffering agents are used other than medium chain fatty acids. In addition, embodiments of the present invention avoid the use of toxic chemicals, such as formaldehyde, which are not incorporated into animal feed, feed ingredients, or pet food. Thus, in one or more embodiments, the chemical buffering agent consists essentially of (or even consists of): one or more medium chain fatty acids. In certain preferred embodiments, the chemical buffering agent consists essentially of (or even consists of): a blend of caproic acid, caprylic acid and capric acid.

Although any effective amount of chemical buffering agent can be used, in one or more embodiments, the chemical buffering agent is introduced into the animal feed (including pet food) or feed ingredient at an inclusion rate of about 0.01 wt.% to about 10 wt.%, preferably about 0.05 wt.% to about 5 wt.%, more preferably about 0.1 wt.% to about 2 wt.%, and most preferably about 0.5 wt.% to about 0.9 wt.%, based on the total weight of the feed or feed ingredient taken as 100 wt.%. Advantageously, the chemical buffering agents described herein, and in particular the medium chain fatty acids, are effective inactivators (i.e., 4-log reductions) of ASFV and CSFV at inclusion rates as low as 0.6 wt% (for ASFV) and 0.5 wt% (for CSFV), which amounts are much lower than the dosages that have been shown to be effective against other microorganisms in the prior art. Thus, in one or more embodiments, the chemical buffer is introduced into the animal feed or feed ingredient at an inclusion rate of less than 2 wt.%, less than 1.5 wt.%, less than 1 wt.%, less than 0.9 wt.%, less than 0.8 wt.%, less than 0.7 wt.%, or less than 0.6 wt.%, based on the total weight of the feed or feed ingredient taken as 100 wt.%. In one or more embodiments, lower doses may also be used, for example when a 4-log reduction of ASFV or CSFV is not required. In one or more such embodiments, the chemical buffering agent can be introduced into an animal feed (including pet food) or feed ingredient at an inclusion rate of about 0.125 wt.% to about 0.5 wt.%, based on the total weight of the feed or feed ingredient taken as 100 wt.%. Thus, in certain other embodiments, the chemical buffering agent is introduced into the animal feed or feed ingredient at an inclusion rate of at least 0.125 wt.%, at least 0.25 wt.%, at least 0.5 wt.%, at least 0.6 wt.%, at least 0.7 wt.%, at least 0.8 wt.%, at least 0.9 wt.%, or at least 1 wt.%, based on the total weight of the feed or feed ingredient taken as 100 wt.%.

In one or more embodiments, sodium bisulfate can be added to the animal feed or feed ingredient in addition to the medium chain fatty acids and/or essential oils. Sodium bisulfate is an acid salt that is considered by the FDA to be "generally regarded as safe" (GRAS) and "natural product". In one or more embodiments, sodium bisulfate can be dissolved in a solution and applied to the surface of an animal food or food ingredient to prevent or reduce bacterial growth. In one or more embodiments, the sodium bisulfate solution is applied to the surface of the dry pet food or pet food ingredient. For example, the solution can be applied to the surface of a dry dog food (roughage) or a dry cat food. In one or more embodiments, the solution can be applied to the surface of the feed or feed ingredient to provide sodium bisulfate at an inclusion rate of about 0.1 wt.% to about 2 wt.%, more preferably about 0.15 wt.% to about 1.5 wt.%, even more preferably about 0.2 wt.% to about 1 wt.%, based on the total weight of the feed or feed ingredient taken as 100 wt.%.

The chemical buffers used according to the present invention can be used to treat a variety of animal feeds or animal feed ingredients. However, in one or more embodiments, the method according to the present invention is particularly suitable for use in swine feed and feed ingredients. In such embodiments, the animal feed or animal feed ingredient may be selected from the group consisting of: complete pig ration, blood meal, pork bone meal (MBM) and spray-dried animal plasma. In one or more embodiments, the animal feed ingredient can include an ingredient selected from the group consisting of: vitamin D, lysine hydrochloride, choline chloride, and soybean meal. In other embodiments, the chemical retardant may be used with pet food and pet food ingredients. In one or more embodiments, the pet food and pet food ingredients comprise dry dog food (roughage) and/or cat food. The term "pet food" means any composition intended for consumption by a pet, and "dry" food generally refers in the art to pet foods having a moisture content of about less than 20% (preferably less than about 15%, more preferably less than about 10%). The term "kibble" is used in the art to refer to particles of dry pet food.

The method according to one or more embodiments of the invention may be used for producing animal or pet feed. Thus, in one embodiment of the invention, there is provided an animal feed or pet food comprising from about 0.01 wt% to about 10 wt%, more preferably from about 0.05 wt% to about 5 wt%, even more preferably from about 0.1 wt% to about 2 wt%, and most preferably from about 0.5 wt% to about 0.9 wt% of a chemical retardant or blend of chemical retardants (such as the chemical retardants and blends described herein), based on the total weight of the feed taken as 100 wt%. In one or more embodiments, an animal feed or pet food is provided that comprises less than 2 wt.%, less than 1.5 wt.%, less than 1 wt.%, less than 0.9 weight%, less than 0.8 wt.%, less than 0.7 wt.%, or less than 0.6 wt.% of a chemical retardant or blend of chemical retardants (e.g., the chemical retardants and blends described herein), based on the total weight of the feed taken as 100 wt.%. In one or more embodiments, an animal feed or pet food is provided that comprises from about 0.125 wt% to about 0.5 wt% of a chemical buffering agent or blend of chemical buffering agents (e.g., the chemical buffering agents and blends described herein) based on the total weight of the feed taken as 100 wt%. In one or more embodiments, an animal feed or pet food is provided that comprises at least 0.5 wt.%, at least 0.6 wt.%, at least 0.7 wt.%, at least 0.8 wt.%, at least 0.9 wt.%, or at least 1 wt.% of a chemical buffering agent or blend of chemical buffering agents (e.g., the chemical buffering agents and blends described herein), based on the total weight of the feed taken as 100 wt.%.

Embodiments of the invention are particularly suitable for use in the transportation, particularly international transportation and storage, of feed and feed ingredients. Prior to the present invention, medium chain fatty acids have not been shown to be effective as a viral-based inhibitor of diseases, particularly viruses, that are foreign to the United states industry. The feed or ingredient may be contaminated with ASFV and/or CSFV during processing. Advantageously, embodiments of the present invention are particularly suitable for inactivating and inhibiting the diffusion of these contaminants. Thus, in one or more embodiments, the method according to the present invention comprises introducing a chemical buffering agent into the feed or feed ingredient after processing. The chemical buffer may be mixed with the feed or feed ingredients for a sufficient time to provide a homogeneous mixture. In one or more embodiments, the methods according to the present invention may prevent or reduce ASFV and/or CSFV in feed and/or ingredients during transport and storage for at least about 90 days after processing, at least about 60 days after processing, at least about 40 days after processing, or at least about 30 days after processing.

Embodiments of the present invention advantageously provide a safe alternative method of preventing or reducing ASFV and/or CSFV in animal or pet feed and ingredients. Existing methods of using hazardous chemicals have shown negative effects on protein and amino acid metabolism in animals. Unlike prior methods, the present invention employs a substantially non-harmful chemical buffer at a dosage found to achieve effective buffering of ASFV and/or CSFV. The chemical buffers used according to the present invention are natural substitutes that pose substantially no risk to the safety of workers or the environment.

Other advantages of various embodiments of the present invention will be apparent to those skilled in the art upon reading the disclosure herein and the following working examples. It will be understood that the various implementations described herein are not necessarily mutually exclusive, unless otherwise indicated herein. For example, features described or depicted in one embodiment may be included in other embodiments as well, but are not necessarily included. Thus, the present invention encompasses various combinations and/or integrations of the specific embodiments described herein.

As used herein, the phrase "and/or," when used in a list of two or more items, means that any one of the listed items can be employed alone or any combination of two or more of the listed items can be employed. For example, if a composition is described as containing or excluding components A, B and/or C, the composition may contain or exclude a alone; b alone; c alone; a combination of A and B; a combination of A and C; a combination of B and C; or a combination of A, B and C.

The present description also uses numerical ranges to quantify certain parameters relating to various embodiments of the invention. It should be understood that when numerical ranges are provided, these ranges are to be construed as providing literal support for claim limitations that only recite the lower value of the range as well as claim limitations that only recite the upper value of the range. For example, a disclosed numerical range of about 10 to about 100 provides literal support for a claim reciting "greater than or equal to about 10" (without an upper limit) and a claim reciting "less than or equal to about 100" (without a lower limit).