Pet food composition

Abstract

Disclosed herein are methods of treating, preventing, or ameliorating periodontal disease in a companion animal, the methods comprising administering to a companion animal in need thereof a composition comprising an effective amount of D-ribose. Also disclosed herein are compositions comprising D-ribose and methods of making the same.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 569,996, filed March 26, 2024, the contents of which are hereby incorporated herein by reference in their entirety. Background Technology

[0003] Many animals, including domestic companion animals (such as canine and feline companion animals), accumulate dental plaque and tartar (dental plaque). These build-ups can lead to periodontal disease. Many pets suffer from gingivitis, which can then progress to periodontitis, eventually leading to even more serious systemic diseases and symptoms. Animals with these dental conditions often also suffer from halitosis and stomatitis. It is estimated that over 70% of dogs have some degree of gingivitis or periodontitis.

[0004] Dental plaque is formed by bacteria colonizing teeth and consists of a layer of bacteria on the erupting surface of teeth and in the gingival folds. More specifically, dental plaque is a biofilm that is typically a pale yellow film that forms naturally on teeth. Like any biofilm, dental plaque is formed by colonizing bacteria that attach themselves to smooth surfaces such as tooth surfaces. Dental plaque containing specific bacterial species can lead to tooth decay and periodontal disease.

[0005] Tartar, or dental plaque, is a recurring calcified deposit on the tooth surface of many animals, including humans, primates, and domesticated dogs and cats. It is generally believed that tartar forms in a continuous process involving plaque accumulation and subsequent calcification of the plaque by saliva, which contains high concentrations of calcium and phosphate. Therefore, tartar is a form of hardened plaque formed by the continuous accumulation of minerals from saliva on plaque already formed on the teeth. The rough surface of tartar provides an ideal medium for further plaque formation in a somewhat autocatalytic manner, which, if left uninterrupted, can threaten gum health.

[0006] Brushing and flossing remove plaque, which forms tartar. However, once tartar has formed, it is often too hard and firmly attached to be removed with a toothbrush. The accumulation of plaque and its subsequent hardening or calcification into tartar or dental plaque eventually leads to irritation and inflammation of the gums, a condition known as gingivitis. Gingivitis can then progress to periodontitis, characterized by the loss of connective tissue fibers from the gums to the teeth and surrounding bone. Tartar (dental plaque) is detrimental to gum health because it acts as a base for increasing plaque formation and retention.

[0007] Since dental plaque is a combination of bacteria, bacterial byproducts that form a glycoprotein matrix, and salivary deposits that bind to the tooth surface, and since the microorganisms that make up dental plaque are considered to be pathogens that cause tartar formation and subsequent inflammation of the oral soft tissues, it is generally believed that measures to inhibit the growth of such microorganisms will inhibit the progression to tartar buildup, gingivitis, and eventually periodontitis.

[0008] Compared to wild animals, pets are more prone to developing plaque and tartar. This is due to the nature of the food pets eat. In the wild, many animals consume food that mechanically erodes plaque and tartar from their teeth. In contrast, pets are typically fed pet food, which usually provides better nutritional value but generally does not subject their teeth to the abrasive forces necessary to clean them.

[0009] Maintaining optimal dental health in pets depends on regular professional cleaning of buildup both above and below the gum line, as well as adequate home care. Mechanical cleaning of the tooth surface through brushing or pet food compositions is an effective means of controlling plaque and tartar buildup in both humans and pets, as these compositions provide mechanical abrasive cleaning when chewed. However, most pet owners are unwilling or unable to provide the routine brushing necessary to maintain their pets' good oral health.

[0010] Therefore, there is a need for food products, compositions, solutions, or methods that can be used to effectively control, reduce, or inhibit the formation of dental plaque and tartar in animals. Summary of the Invention

[0011] This summary is intended only to provide a simplified overview of some aspects of one or more embodiments of the present disclosure. Other applicable areas of the disclosure will become apparent from the detailed description provided below. This summary is not an exhaustive overview, nor is it intended to identify key or essential elements of the teachings, nor is it intended to define the scope of the disclosure. Rather, its purpose is merely to present one or more concepts in a simplified form as a prelude to the following detailed description.

[0012] Various aspects of this disclosure relate to pet food compositions and methods of using them for reducing plaque and tartar buildup and / or improving oral health in pets. According to one aspect, a composition, such as a pet food composition, is provided comprising D-ribose.

[0013] According to some aspects, a method for treating, preventing, or alleviating periodontal disease in companion animals is provided, the method comprising administering to the companion animal in need a composition comprising an effective amount of D-ribose (e.g., a pet food composition). In some embodiments, the D-ribose is present in the composition in an amount ranging from about 1 wt% to about 5 wt%, preferably from about 2 wt% to about 4 wt%, or about 2 wt%, based on the total weight of the composition. The method may include providing and / or feeding the canine with the composition disclosed herein. In some cases, the method may include providing and / or feeding the canine with the composition for 1 day or more, preferably 5 days or more, preferably 7 days or more, preferably 10 days or more, preferably 14 days or more, preferably 30 days or more, or preferably 42 days or more.

[0014] According to some embodiments, periodontal disease in a companion animal is treated or alleviated by reducing the amount of at least one of tartar or plaque on the companion animal's teeth compared to before the composition is applied to the companion animal, and in some embodiments, the methods disclosed herein reduce at least one of Fusobacterium nucleatum or Porphyromonas gingivalis present on the teeth of the companion animal compared to before the composition is applied to the companion animal.

[0015] In various embodiments, the composition is a pet food composition and is in the form of dry extruded food, wet cakes, dental chews, or treats, preferably dry extruded food, and in some embodiments, the D-ribose is in the form of a coating covering the dry extruded food. In various embodiments, the composition is in the form of a dental cleaning agent for oral care, such as a paste, gel, mouthwash, dental rinse, or tablet (e.g., a toothpaste composition).

[0016] According to some embodiments, the pet food composition further comprises at least one mechanical component for cleaning teeth, such as a transverse fibrous striation matrix.

[0017] This document also discloses a composition (e.g., a pet food composition) for treating, preventing, or alleviating periodontal disease in companion animals, the composition comprising an effective amount of D-ribose. Optionally, the composition is a pet food composition and is in the form of extruded food containing a coating comprising the D-ribose, or alternatively, the composition is a dental floss and is in the form of a paste, gel, or tablet (e.g., a toothpaste composition). In some embodiments, the composition does not contain any additional sugar. In some embodiments, the composition is a coated pet food composition, and the coating further comprises at least one palatability enhancer.

[0018] This document further discloses a method for reducing dental plaque buildup, the method comprising administering a composition as disclosed herein to a companion animal. In another embodiment, this document discloses a method for reducing dental plaque buildup, the method comprising administering a composition as disclosed herein to a companion animal. In yet another embodiment, a method for reducing the amount of Fusobacterium nucleatum and / or Porphyromonas gingivalis in the oral cavity is provided, the method comprising administering a composition as disclosed herein to a companion animal.

[0019] According to another aspect, this document discloses a method for preparing a pet food composition for treating, preventing, or alleviating symptoms of periodontal disease, the method comprising: (a) mixing wet and dry ingredients to form a dough; (b) extruding the dough to form an extruded puffed food; (c) drying the extruded puffed food; and (d) coating the dried puffed food with a localized liquid ingredient and / or dry ingredient, wherein D-ribose is applied at step (a) and / or step (d) in an amount effective in treating, preventing, or alleviating symptoms of periodontal disease in a companion animal consuming the pet food composition. In some embodiments, the D-ribose is added to the puffed food at step (a) when the wet and dry ingredients are mixed to form a dough, and in some embodiments, the D-ribose is added to the puffed food at step (d) as the localized liquid ingredient and / or dry ingredient is applied. In some embodiments of the method for preparing a pet food composition disclosed herein, the D-ribose is applied in an amount ranging from about 1 wt% to about 5 wt%, preferably from about 2 wt% to about 4 wt% or about 2 wt%, based on the total weight of the pet food composition.

[0020] According to certain embodiments of any aspect of this disclosure, the companion animal is a cat or a dog, preferably a dog. Attached Figure Description

[0021] The features and advantages of this disclosure will become apparent from the following more detailed description of certain embodiments of the disclosure and from the accompanying drawings, in which:

[0022] Figure 1 This is a schematic diagram illustrating the experimental design of a food study as described in Example 1 of this article, where “A” represents the positive control food, “B” represents test food 1, “C” represents the negative control food, and “D” represents test food 2.

[0023] Figure 2 This is a bar graph of the full dental plaque scores of dogs fed negative control food, positive control food, test food 1, and test food 2. The letters above the bars indicate significantly different (p < 0.05) results obtained through mixed model analysis and subsequent Tukey post-hoc test as described in Example 1.

[0024] Figure 3 The bar graph shows the abundance levels of Fusobacterium nucleatum in subgingival plaque samples from healthy and unhealthy teeth (left) and subgingival plaque samples from dogs fed negative control food, positive control food, test food 1, and test food 2 (right). The letters above the bars indicate significantly different (p < 0.05) results obtained through mixed model analysis and subsequent post-hoc graph base test as described in Example 1.

[0025] Figure 4 This is a bar graph showing the abundance levels of Porphyromonas gingivalis in subgingival plaque samples from healthy and unhealthy teeth (left) and in subgingival plaque samples from dogs fed negative control food, positive control food, test food 1, and test food 2 (right). The letters above the bars indicate significantly different (p < 0.05) results obtained through mixed model analysis and subsequent post-hoc graph base test as described in Example 1.

[0026] Figure 5 This is a graph showing the distribution of gingivitis scores and dental plaque scores for subgingival plaque samples collected as described in Example 1, where lower scores indicate healthier teeth.

[0027] Figure 6 This is a graph showing a linear regression analysis of Fusobacterium nucleatum using dental plaque score (left), dental tartar score (middle), and dental gingivitis score (right) as described in Example 1, where lower scores indicate healthier teeth, and RSME indicates root mean square error.

[0028] Figure 7This is a graph showing a linear regression analysis of Porphyromonas gingivalis using dental plaque score (left), dental tartar score (middle), and dental gingivitis score (right) as described in Example 1, where lower scores indicate healthier teeth, and RSME indicates root mean square error.

[0029] Figure 8 This is a graph showing a linear regression analysis of Fusobacterium nucleatum using full-mouth plaque score (left), full-mouth tartar score (middle), and full-mouth gingivitis score (right) as described in Example 1, where lower scores indicate healthier teeth, and RSME indicates root mean square error.

[0030] Figure 9 This is a graph showing a linear regression analysis of Porphyromonas gingivalis using full-mouth plaque score (left), full-mouth tartar score (middle), and full-mouth gingivitis score (right) as described in Example 1, where lower scores indicate healthier teeth, and RSME indicates root mean square error.

[0031] It should be understood that the various aspects of this disclosure are not limited to the compositions, arrangements, and tools shown in the accompanying drawings. Detailed Implementation

[0032] For illustrative purposes, the principles of this disclosure are described with reference to various exemplary embodiments. While certain embodiments of this disclosure have been specifically described herein, those skilled in the art will readily recognize that the same principles apply equally to and can be employed in other compositions and methods. Before explaining the disclosed embodiments in detail, it should be understood that this disclosure is not necessarily limited to the details of any particular embodiment disclosed herein. The terminology used herein is for descriptive purposes and not for limitation.

[0033] As used herein and in the appended claims, the singular forms “a / an” and “the” include plural references unless the context clearly indicates otherwise. The singular form of any class of ingredients refers not only to one ingredient within that class but also to mixtures of those ingredients. The terms “a / an,” “one or more,” and “at least one” are used interchangeably herein. The terms “comprising,” “including,” and “having” are used interchangeably. The term “include” should be interpreted as “including, but not limited to.” The term “including” should be interpreted as “including, but not limited to.”

[0034] As used throughout, a range is used as a concise way to describe each value within a range and every single value. Any value within the range can be chosen as the endpoint of the range. Therefore, the range 1 to 5 specifically includes 1, 2, 3, 4, and 5, as well as subranges such as 2 to 5, 3 to 5, 2 to 3, 2 to 4, 1 to 4, etc. When referring to numbers, the term "about" means any number within 10% of the range stated. For example, the phrase "about 2 wt.%" refers to a number between and including 1.8 wt.% and 2.2 wt.%.

[0035] All references cited herein are incorporated herein by reference in their entirety. In the event of any conflict between the definitions in this disclosure and those in the cited references, this disclosure shall prevail.

[0036] Unless otherwise specified, abbreviations and symbols used herein have their usual meanings. The abbreviation “wt.%” indicates a weight percentage relative to the pet food composition. The symbol “°” refers to a degree, such as temperature or angle. The symbols “h,” “min,” “mL,” “nm,” and “μm” refer to hours, minutes, milliliters, nanometers, and micrometers, respectively. The abbreviation “UV-VIS,” referring to a spectrometer or spectroscopy method, means ultraviolet-visible. The abbreviation “rpm” refers to revolutions per minute.

[0037] Any member of the list of species used to exemplify or define a genus may be different from, overlap with, be a subset of, be equivalent to, or be nearly identical to, any other member of the list of species. Furthermore, unless explicitly stated otherwise, as in the description of the Markush Formation, the list of species used to define or exemplify a genus is open, provided that other species may exist that define or exemplify the genus as well as or better than any of the other species listed.

[0038] All components and elements actively described in this disclosure may be excluded negatively from the claims. In other words, the pet food compositions of this disclosure may be free of or substantially free of all components and elements positively described throughout this disclosure. In some cases, the compositions of this disclosure may be substantially free of non-accidental amounts of the ingredients or compounds described herein. A non-accidental amount of an ingredient or compound is the amount of said ingredient or compound added alone to the composition. For example, the composition may be substantially free of non-accidental amounts of an ingredient or compound, although such an ingredient or compound may be present as a component of a blend of two or more compounds included as a raw material. Unless otherwise defined or described herein, “substantially free” generally means an amount of an ingredient or compound based on the total weight of the composition on a dry matter basis of about 2 wt.% or less, about 1.5 wt.% or less, about 1 wt.% or less, about 0.5 wt.% or less, about 0.1 wt.% or less, or about 0.05 wt.% or less, about 0.01 wt.% or less, or about 0.001 wt.% or less.

[0039] Some components among the various categories of components identified may overlap. In such cases where overlap is possible and the composition comprises two components (or the composition comprises more than two overlapping components), the overlapping compounds do not represent more than one component. For example, certain components or ingredients may be characterized as both archaea and amaranth. If a particular composition describes both archaea and amaranth, then a compound characterized as both archaea and amaranth will act as only one of archaea or amaranth, not both.

[0040] As used herein, the term "pet" is used interchangeably with "companion animal" and refers to any species of animal kept as a pet by a caregiver, or any animal of various species that has been widely domesticated as a pet, including canines (Dog (Canis familiaris)) and felines (Dog (Felis domesticus)). Therefore, pets can include, but are not limited to, working dogs, pet dogs, cats kept for rodent control (i.e., farm cats), pet cats, ferrets, birds, reptiles, rabbits, and fish.

[0041] In the context of water / moisture content in food and food ingredients, dry matter represents all substances in a sample other than water, including, for example, proteins, fiber, fats, carbohydrates, minerals, etc. Dry matter weight is the total weight minus the weight of any water. Those skilled in the art will readily recognize and understand nutrient content and percentages expressed as dry matter mass, dry matter weight, and dry matter percentage.

[0042] Daily dry matter intake is calculated as total daily nutrient intake excluding all water. For example, the amount of a component equivalent to a specific percentage of daily nutrient intake refers to the amount of said component in dry matter form (i.e., excluding all water) relative to the total amount of dry matter (also excluding all water) consumed during the day.

[0043] "Daily nutrient intake" and "Total daily nutrient intake" refer to daily dry matter intake. That is, the weight of water is not included when calculating the amount of nutrients consumed each day. To calculate the percentage of total daily intake in dry matter terms, water is removed from the total intake to obtain the total daily dry matter intake, and the percentage of the component is based on the amount of the component present in dry matter form.

[0044] As used herein, “ingredient” means any component of a composition. The term “nutrient” refers to a substance that provides nutrition and thus has nutritional value. In some cases, an ingredient may contain more than one “nutrient”; for example, a composition may contain corn, which contains important nutrients including both protein and carbohydrates.

[0045] As used herein, the term "effective amount" refers to an amount of a compound, material, or composition as described herein that is effective in achieving a specific biological outcome, such as the treatment, prevention, or relief of periodontal disease, including conditions selected from plaque accumulation, tartar, tooth discoloration, gingivitis, periodontitis, stomatitis, halitosis, and combinations thereof. Such outcomes can be achieved, for example, by feeding animals requiring such treatment with the compositions disclosed herein.

[0046] As used herein, the term "coating" means a partial or complete covering of at least a portion of a surface covering the matrix of an ingredient (e.g., the surface of an extruded grain). In one instance, the extruded grain may be partially coated such that only a portion of the surface of the extruded grain is covered, and in another instance, the extruded grain may be completely coated.

[0047] As used herein, the term "pet food composition" refers to a composition intended for consumption by animals (e.g., dogs or cats). Such foods can be formulated into nutritionally balanced compositions suitable for daily feeding and can be prepared as dry food compositions, such as extruded food, or as dietary supplements or snacks that may not necessarily be nutritionally balanced. As used herein, the term "extruded food" includes particulate pellet components of animal food with a texture that can vary from hard to soft and can have an internal structure ranging from an expanded structure to a dense structure. Extruded food can be formed through an extrusion process.

[0048] As used herein, the term "dental cleanser" refers to a paste, gel, liquid, or solid composition for oral care (such as oral care for companion animals). In some embodiments, a dental cleanser composition may be a paste, gel, or a combination of a paste and a gel. In some embodiments, a dental cleanser composition is toothpaste, and in some embodiments, a dental cleanser composition is mouthwash or dental rinse.

[0049] This disclosure relates to compositions (including pet food compositions and dental cleaning agents), methods of use thereof for treating, preventing or alleviating periodontal disease, and methods of reducing plaque and tartar buildup and / or improving oral health in companion animals. In embodiments of this disclosure, the compositions disclosed herein contain D-ribose.

[0050] D-ribose is a monosaccharide and a fundamental component of adenosine triphosphate (ATP), which provides energy to host somatic cells. Additionally, D-ribose acts as a quorum sensing inhibitor. Specifically, D-ribose inhibits autoinducer 2, a universal quorum sensing molecule that mediates intergeneric signaling in multi-species bacterial communities to induce and maintain biofilm formation. In in vitro biofilm assays and co-coagulation cultures of both *Fusobacterium nucleatum* and *Porphyromonas gingivalis*, D-ribose has shown inhibitory properties against the quorum sensing molecule autoinducer 2. Therefore, without being bound by theory, it is believed that D-ribose present in the compositions disclosed herein acts by modulating potentially pathogenic bacterial species to inhibit and / or reduce biofilm formation, thereby preventing periodontal diseases such as gingivitis and periodontitis.

[0051] Bacteria play a crucial role in the etiology of periodontal disease through the accumulation of dental plaque biofilms, and the accumulation of specific pathogenic bacterial species is key to activating the host's inflammatory response. As plaque buildup extends from the gingival margin to the subgingival sulcus, the anaerobic environment facilitates the proliferation of certain anaerobic bacterial species, including pathogenic species that act as late-term colonizers. The persistent presence of these late-term colonizers can lead to inflammation of the gingival tissue (gingivitis) and induce an immune response in the host. Regularly disrupting the plaque buildup and maturation process can help prevent gingivitis and periodontitis. This regular disruption can be achieved through mechanical means (such as brushing, scaling, dental chewing agents, and fiber-based techniques) and / or chemical means (such as through polyphosphates, antibiotics, and biofilm-degrading agents). Furthermore, prevention using specific ingredients (e.g., D-ribose) that target late-term colonizers may help treat, prevent, and / or alleviate the symptoms of periodontal disease.

[0052] Late-term colonizing bacteria can be grouped into the "Orange Complex" and the "Red Complex" bacterial species. The "Orange Complex" includes three species: *Fusobacterium nucleatum*, *Campylobacter rectus*, and *Prevotella intermedia*. *Fusobacterium nucleatum* is believed to play a role in halitosis, pulpitis, and potential oral cancer, and is considered a major species in the development of periodontitis due to creating favorable conditions that preferentially reduce the efficiency of host neutrophils. *Fusobacterium nucleatum* also acts as a "bridging species," metabolically linking with the "Red Complex" species. One such species in the “red complex” is *Porphyromonas gingivalis*, a major pathogenic agent found in subgingival plaque in patients with periodontal disease. Therefore, interfering with the persistence of *Fusobacterium nucleatum*, for example by using compositions containing D-ribose as disclosed herein, could help limit the establishment of *Porphyromonas gingivalis*, thereby treating, preventing, and / or alleviating symptoms of various periodontal diseases, including gingivitis.

[0053] Therefore, this document discloses a composition comprising D-ribose. In some embodiments, the composition (e.g., a pet food composition or a dental cleaning agent) may contain an amount of D-ribose that effectively treats, prevents, and / or alleviates symptoms of various periodontal diseases, including gingivitis. In some embodiments, the compositions disclosed herein comprise D-ribose in amounts ranging from about 0.5 wt.% to about 10 wt.%, such as about 0.5 wt.% to about 5 wt.%, about 0.5 wt.% to about 4.5 wt.%, about 0.5 wt.% to about 4 wt.%, about 0.5 wt.% to about 3.5 wt.%, about 0.5 wt.% to about 3 wt.%, about 0.5 wt.% to about 2.5 wt.%, or about 0.5 wt.% to about 2 wt.%; about 1 wt.% to about 5 wt.%, about 1 wt.% to about 4.5 wt.%, about 1 wt.% to about 4 wt.%, about 1 wt.% to about 3.5 wt.%, about 1 wt.% to about 3 wt.%, about 1 wt.% to about 2.5 wt.%, or about 1 wt.% to about 2 wt.%; about 1.5 wt.% to about 5 wt.%. wt.%, about 1.5 wt.% to about 4.5 wt.%, about 1.5 wt.% to about 4 wt.%, about 1.5 wt.% to about 3.5 wt.%, about 1.5 wt.% to about 3 wt.%, about 1.5 wt.% to about 2.5 wt.% or about 1.5 wt.% to about 2 wt.%; or about 2 wt.%.

[0054] In some embodiments, the composition is formulated to contain only D-ribose as the sole added sugar. For example, in some embodiments, the composition contains no or substantially no other added sugars, such as dextrose, glucose, galactose, fructose, sucrose, lactose, maltose, arabinose, xylose, mannose, erythrose, and / or threose.

[0055] Depending on the specific combination of other ingredients and / or the form of the composition, the formulation of the compositions disclosed herein may include or exclude suitable components (as listed below). In some embodiments, the pet food compositions disclosed herein may be in the form of stand-alone pet food, as a supplement to pet food, as a pet food treat, or in similar forms. In some embodiments, the pet food compositions disclosed herein may contain protein, carbohydrates, fat, and fiber.

[0056] In some embodiments, the pet food compositions of this disclosure include proteins. The proteins can be provided from any of a variety of sources known to those skilled in the art, including plant sources, animal sources, microbial sources, or combinations thereof. For example, animal sources can include meat, meat by-products, seafood, dairy products, eggs, etc. Meat, for example, can include animal meat such as poultry, fish, and mammals including cattle, pigs, sheep, goats, etc. Meat by-products can include, for example, lungs, kidneys, brain, liver, stomach, and intestines. Plant proteins include, for example, soybeans, cottonseed, and peanuts. Microbial sources can be used to synthesize amino acids (e.g., lysine, threonine, tryptophan, methionine) or complete proteins (such as proteins from sources listed herein).

[0057] Proteins disclosed herein in pet food compositions may be present in a quantity that can vary. For example, based on the total weight of the pet food composition on a dry matter basis, the pet food composition may include protein in an amount ranging from about 15 wt.% to about 55 wt.%. In some embodiments, based on the total weight of the pet food composition on a dry matter basis, the amount of protein present in the pet food composition is about 15 wt.% to about 50 wt.%, about 15 wt.% to about 45 wt.%, about 15 wt.% to about 40 wt.%, about 15 wt.% to about 35 wt.%, about 15 wt.% to about 30 wt.%; about 20 wt.% to about 55 wt.%, about 20 wt.% to about 50 wt.%, about 20 wt.% to about 45 wt.%, about 20 wt.% to about 40 wt.%, about 20 wt.% to about 35 wt.%, about 20 wt.% to about 30 wt.%; about 23 wt.% to about 55 wt.%, about 23 wt.% to about 50 wt.%, about 23 wt.% to about 45 wt.%, about 23 wt.% to about 40 wt.%, about 23 wt.% to about 35 wt.%. wt.%; about 26 wt.% to about 55 wt.%, about 26 wt.% to about 50 wt.%, about 26 wt.% to about 45 wt.%, about 26 wt.% to about 40 wt.%, about 26 wt.% to about 35 wt.%; about 29 wt.% to about 55 wt.%, about 29 wt.% to about 50 wt.%, about 29 wt.% to about 45 wt.%, about 29 wt.% to about 40 wt.%, about 29 wt.% to about 35 wt.%; about 32 wt.% to about 55 wt.%, about 32 wt.% to about 50 wt.%, about 32 wt.% to about 45 wt.%, about 32 wt.% to about 40 wt.%; about 35 wt.% to about 55 wt.%, about 35 wt.% to about 50 wt.%, about 35 wt.% to about 45 wt.%, about 35 wt.% to about 40 wt.%; about 38 wt.% to about 55 wt.%, about 38 wt.% to about 50 wt.%, about 38 wt.% to about 45 wt.%; about 41 wt.% to about 55 wt.%, about 41 wt.% to about 50 wt.%; or about 43 wt.% to about 55 wt.%, including all ranges and subranges thereof.

[0058] Examples of proteins or protein components may include chicken meal, chicken meat, chicken by-product meals, lamb, lamb meal, turkey, turkey meal, beef, beef by-products, offal, fish meal, sausage, kangaroo, white fish, venison, soy meal, soy protein isolate, soy protein concentrate, corn gluten meal, corn protein concentrate, dried distillers grains and / or dried distillers grains solubles, and single-cell proteins such as yeast, algae, and / or bacterial cultures.

[0059] The pet food compositions disclosed herein may also contain free amino acids, peptides, and combinations thereof. Amino acids may be selected from tryptophan, taurine, histidine, carnitine, carnosine, alanine, cysteine, arginine, methionine (including DL-methionine and L-methionine), tryptophan, lysine, asparagine, aspartate, phenylalanine, valine, threonine, isoleucine, histidine, leucine, glycine, glutamine, taurine, tyrosine, homocysteine, ornithine, citrulline, glutamate, proline, and / or serine, or combinations of two or more. In some cases, one or more amino acids may contain essential amino acids. Essential amino acids are amino acids that cannot be synthesized de novo or by the organism in sufficient quantities, and therefore must be provided in the diet. Essential amino acids vary between species, depending on the organism's metabolism. For example, the essential amino acids for dogs and cats (and humans) are generally considered to be phenylalanine, leucine, methionine, lysine, isoleucine, valine, threonine, tryptophan, histidine, and arginine. Additionally, taurine, although technically a derivative of cysteine ​​rather than an amino acid, is an essential nutrient for cats.

[0060] The pet food compositions disclosed herein may further comprise one or more sources of fat and / or carbohydrates, including those known in the art. Illustrative examples of suitable sources of carbohydrates and fats include, but are not limited to, animal fats, fish oils, vegetable oils, meat, meat by-products, grains, other animal or plant sources and mixtures thereof. Grains include wheat, sorghum, corn, barley, rice, etc.

[0061] In some embodiments, the pet food composition may contain about 70 wt.% or less of carbohydrates based on the total weight of the pet food composition on a dry matter basis. In some embodiments, the amount of carbohydrates in the pet food composition is in the range of about 5 wt.% to about 65 wt.%, such as about 25 wt.% to about 50 wt.%, about 30 wt.% to about 50 wt.%, about 35 wt.% to about 50 wt.%, about 40 wt.% to about 50 wt.%, about 45 wt.% to about 50 wt.%, or about 47 wt.% to about 50 wt.%.

[0062] As used herein, the term "carbohydrate" includes polysaccharides (e.g., starch and dextrin) and sugars (e.g., sucrose, lactose, maltose, glucose, and fructose) that are metabolized into energy upon hydrolysis. Typically, the percentage of carbohydrates can be calculated as nitrogen-free extract ("NFE"), which can be calculated as follows: NFE% = 100% - (moisture%) - (protein%) - (fat%) - (ash%) - (crude fiber%). Those skilled in the art can manipulate the texture of the final product by appropriately balancing the sources of carbohydrates. For example, short-chain polysaccharides tend to be sticky and gelatinous, while long-chain polysaccharides tend to be less sticky and less gelatinous compared to shorter-chain polysaccharides. Therefore, a desired texture for pet food can be achieved by adding the desired amount of longer-chain polysaccharides and modified starches (such as natural or modified starch, cellulose, etc.). The carbohydrate mixture may additionally contain optional components such as added salts, spices, flavorings, vitamins, minerals, taste agents, colorings, etc. The amount of optional components may depend at least in part on the nutritional requirements of different life stages of the animal.

[0063] Carbohydrates can be supplied from any of a variety of sources known to those skilled in the art, including but not limited to oat fiber, cellulose, peanut shells, beet pulp, parboiled rice, corn starch, corn gluten meal, grain flour, and sorghum. Cereals supplying carbohydrates can include, but are not limited to, wheat, durum wheat, wheat grits, corn, barley, and rice. In some embodiments, the carbohydrate component comprises a mixture of one or more carbohydrate sources. The carbohydrate content of a food can be determined by many methods known to those skilled in the art.

[0064] In some cases, the fat in the pet food composition is crude fat. Based on the total weight of the pet food composition on a dry matter basis, crude fat may be included in the composition in any amount known in the art (e.g., from about 2 wt.% to about 50 wt.%). In some embodiments, based on the total weight of the pet food composition on a dry matter basis, the pet food composition contains approximately 2 wt.% to about 40 wt.%, approximately 2 wt.% to about 35 wt.%, approximately 2 wt.% to about 30 wt.%, approximately 2 wt.% to about 25 wt.%, approximately 2 wt.% to about 20 wt.%, approximately 2 wt.% to about 15 wt.%, approximately 5 wt.% to about 35 wt.%, approximately 5 wt.% to about 30 wt.%, approximately 5 wt.% to about 25 wt.%, approximately 5 wt.% to about 20 wt.%, approximately 5 wt.% to about 15 wt.%, approximately 10 wt.% to about 35 wt.%, approximately 10 wt.% to about 30 wt.%, approximately 10 wt.% to about 25 wt.%, approximately 10 wt.% to about 20 wt.%, approximately 10 wt.% to about 15 wt.%, or approximately 11 wt.% to about 15 wt.%. Crude fat percentage (wt.%). In some cases, it may be preferred to obtain about 50 wt.% or more, about 60 wt.% or more, about 70 wt.% or more, about 80 wt.% or more, or about 90 wt.% or more total fat from animal sources. Alternatively, about 50 wt.% or more, about 60 wt.% or more, about 70 wt.% or more, about 80 wt.% or more, or about 90 wt.% or more total fat from plant sources.

[0065] The pet food compositions disclosed herein may also contain one or more types of fiber, which may be classified as soluble or insoluble fiber. Illustrative but non-limiting sources of fiber include those from various sources, such as plant fiber sources, like cellulose, beet pulp, peanut shells, and soybean fiber.

[0066] Based on the total weight of the pet food composition on a dry matter basis, the pet food composition disclosed herein can be formulated to include fiber in amounts ranging from about 2 wt.% to about 50 wt.%. For example, based on the total weight of the pet food composition on a dry matter basis, the amount of fiber present in the pet food composition may be about 2 wt.% to about 45 wt.%, about 2 wt.% to about 40 wt.%, about 2 wt.% to about 35 wt.%, about 2 wt.% to about 20 wt.%, about 5 wt.% to about 45 wt.%, about 5 wt.% to about 40 wt.%; about 5 wt.% to about 35 wt.%, about 5 wt.% to about 30 wt.%, about 5 wt.% to about 25 wt.%, about 5 wt.% to about 20 wt.%, about 8 wt.% to about 45 wt.%, about 8 wt.% to about 40 wt.%, about 8 wt.% to about 35 wt.%; about 8 wt.% to about 30 wt.%, about 8 wt.% to about 25 wt.%, about 8 wt.% to about 20 wt.% or about 8 wt.% to about 17 wt.%. wt.%, including its range and subranges.

[0067] Dietary fiber refers to the components of plants that resist digestion by digestive enzymes in animals. Dietary fiber includes soluble and insoluble fiber. Soluble fiber resists digestion and absorption in the small intestine and undergoes complete or partial fermentation in the large intestine; examples include beet pulp, guar gum, chicory root, psyllium husk, pectin, blueberries, cranberries, squash, apples, oats, legumes, citrus fruits, barley, or peas. Insoluble fiber can be provided from a variety of sources, including, for example, cellulose, whole-wheat products, wheat and oats, corn bran, flaxseed, grapes, celery, green beans, cauliflower, potato peels, fruit peels (e.g., pear peels), vegetable peels, peanut shells, almond shells, walnut shells, pecan shells, citrus pulp, beet pulp, and soy fiber. In some embodiments, dietary fiber may be selected from pecan shells, citrus pulp, beet pulp, and combinations of two or more thereof. Crude fiber includes the indigestible components contained in the cell walls and cell contents of plants such as grains, for example, the husks of grains such as rice, corn, and legumes.

[0068] In some embodiments, the pet food compositions of this disclosure may also contain nutritional balancers, such as vitamin and mineral supplements, and food ingredients, available from various sources known to those skilled in the art. Vitamins and minerals are included in amounts necessary to avoid deficiency and maintain health. These amounts are readily available in the art. The Association of American Feed Control Officials (AAFCO) provides recommended amounts of such nutrients for dogs and cats. See the Association of American Feed Control Officials, Inc., Official publication, pp. 159-162 (2011). Vitamins commonly used as food additives include vitamin A, vitamin B1, vitamin B2, vitamin B6, and vitamin B6. 12 Vitamins D, biotin, vitamin K, folic acid, inositol, niacin, and pantothenic acid. Minerals and trace elements that can be used as food additives include calcium, phosphorus, sodium, potassium, magnesium, copper, zinc, chloride, iron, selenium, iodine, and iron.

[0069] In some other embodiments, in addition to vitamins and minerals, the pet food compositions of this disclosure may contain other ingredients such as fillers, palatability enhancers, binders, flavoring agents, stabilizers, emulsifiers, sweeteners, colorants, buffers, salts, coatings, etc., known to those skilled in the art. Stabilizers include substances that tend to increase the shelf life of the composition, such as preservatives, synergists and complexing agents, packaging gases, stabilizers, emulsifiers, thickeners, gelling agents, and humectants. Examples of emulsifiers and / or thickeners include gelatin, cellulose ethers, starch, starch esters, starch ethers, and modified starches. The specific amounts of each composition component, food ingredient, and other ingredient will depend on a variety of factors, such as the specific components and ingredients included in the composition; the species of the animal; the animal's age, weight, overall health condition, sex, and diet; the animal's consumption rate; the type of disease or symptom being treated; and so on. Therefore, the amounts of components and ingredients can vary widely and may deviate from the preferred proportions described herein.

[0070] In some embodiments, the pet food composition contains moisture. Moisture may be present in varying amounts or concentrations. In one embodiment, moisture may be present in amounts from about 3 wt.% to about 20 wt.% based on the total weight of the pet food composition. For example, moisture may be present in amounts from about 3 wt.%, about 5 wt.%, about 5.5 wt.%, about 6 wt.%, about 6.5 wt.%, about 7 wt.%, about 7.5 wt.%, about 8 wt.%, about 8.5 wt.%, about 9 wt.%, about 9.5 wt.%, about 10 wt.%, about 10.5 wt.%, about 11 wt.%, about 11.5 wt.%, about 12 wt.%, about 12.5 wt.%, about 13 wt.%, about 13.5 wt.%, about 14 wt.%, about 14.5 wt.%, or about 15 wt.% based on the total weight of the pet food composition. In another embodiment, moisture may be present in amounts of about 6 wt.% to about 12 wt.%, about 9 wt.% to about 13 wt.%, about 9 wt.% to about 11 wt.%, or about 9 wt.% to about 13 wt.% based on the total weight of the pet food composition. In some embodiments, moisture may be present in amounts of about 5 wt.% to about 12 wt.%, about 6 wt.% to about 11 wt.%, or about 7 wt.% to about 10 wt.% based on the total weight of the pet food composition. In other embodiments, moisture may be present in amounts of about 65 wt.% to about 85 wt.%, about 60 wt.% to about 80 wt.%, or about 60 wt.% to about 75 wt.% based on the total weight of the pet food composition.

[0071] The pet food compositions disclosed herein can be wet or dry compositions, and ingredients including, for example, D-ribose can be incorporated into the food composition and / or incorporated onto the surface of any composition component, for example, by spraying, agglomerating, dusting, or depositing onto a surface. Additionally, pet food compositions can be formulated and manufactured in various forms and / or consistencies. For example, pet food compositions can be, for example, dry, moist, or semi-moist animal food compositions. "Semi-moist" refers to a food composition containing about 25% to about 35% moisture. "Moist" food refers to a food composition with a moisture content of about 60% to 90% or higher. "Dry" food refers to a food composition having a moisture content of about 3% to about 12% and is typically manufactured in the form of small pieces or extruded food.

[0072] Food products may also include components of more than one consistency, such as soft, chewable meat-like pellets or chunks, and extruded food having an outer coating and an internal "core" component. In some embodiments, pet food compositions may be in the form of extruded food or food extruded food. In some embodiments, the moisture or water content of food extruded food is less than 15% by weight. The texture of food extruded food can vary from hard to soft. The internal structure of food extruded food can vary from expanded to dense. Food extruded food can be formed by an extrusion process or a baking process. In non-limiting examples, food extruded food may have a uniform internal structure or a varied internal structure. For example, food extruded food may include a core and a coating to form a coated extruded food. It should be understood that when the terms "extruded food" or "food extruded food" are used, they may refer to uncoated extruded food or coated extruded food.

[0073] The compositions disclosed herein may additionally contain other additives in amounts and combinations familiar to those skilled in the art. Such additives should be present in amounts that do not impair the purposes and effects provided by the invention. Examples of additives include substances with stabilizing effects, sensory substances, processing aids, and substances that provide nutritional benefits.

[0074] For example, stabilizing substances may include substances that tend to increase the shelf life of pet food compositions. Other examples of such additives potentially suitable for inclusion in the compositions of the present invention include, for example, preservatives, antioxidants, synergists and complexing agents, packaging gases, stabilizers, emulsifiers, thickeners, gelling agents, and humectants. Examples of emulsifiers and / or thickeners include gelatin, cellulose ethers, starch, starch esters, starch ethers, and modified starches. Additives for coloring, palatability, and nutritional purposes may include colorants, salts (including but not limited to sodium chloride, potassium citrate, potassium chloride, and other edible salts), vitamins, minerals, and flavorings. Other additives may include glucosamine, chondroitin sulfate, vegetable extracts, herbal extracts, etc.

[0075] Based on the total weight of the pet food composition on a dry matter basis, the amount of such additives in the pet food composition may be up to about 5 wt.%. For example, based on the total weight of the pet food composition on a dry matter basis, the additive may be in the following proportions: about 0.01 wt.% to about 5 wt.%, about 0.01 wt.% to about 4 wt.%, about 0.01 wt.% to about 4 wt.%, about 0.01 wt.% to about 3 wt.%, about 0.01 wt.% to about 2 wt.%, about 0.01 wt.% to about 1 wt.%; about 0.1 wt.% to about 5 wt.%, about 0.1 wt.% to about 4 wt.%, about 0.1 wt.% to about 4 wt.%, about 0.1 wt.% to about 3 wt.%, about 0.1 wt.% to about 2 wt.%, about 0.1 wt.% to about 1 wt.%; about 1 wt.% to about 5 wt.%, about 1 wt.% to about 4 wt.%, about 1 wt.% to about 4 wt.%, about 1 wt.% to about 3 wt.%, about 1 wt.% to about 2 wt.%. wt.%; about 2 wt.% to about 5 wt.%, about 2 wt.% to about 4 wt.%, about 2 wt.% to about 4 wt.%, about 2 wt.% to about 3 wt.%; about 3 wt.% to about 5 wt.%, about 3 wt.% to about 4 wt.%; or about 4 wt.% to about 5 wt.% are present.

[0076] In certain embodiments, pet food compositions and food products are formulated to address specific nutritional differences between animal species and breeds, as well as among a number of attributes of the animal. For example, cat food is typically formulated based on life stage, age, size, weight, body composition, and breed.

[0077] Pet food compositions may include, or in some cases exclude, one or more ingredients selected from the following: beef broth, brewer's dry yeast, eggs, egg products, flaxseed meal, DL-methionine, amino acids, leucine, lysine, arginine, cysteine, cystine, aspartic acid, polyphosphates, sodium pyrophosphate, sodium tripolyphosphate; zinc chloride, copper gluconate, stannous chloride, stannous fluoride, sodium fluoride, triclosan, glucosamine hydrochloride, chondroitin sulfate, green-lipped mussels, blue-lipped mussels, methanesulfonylmethane (MSM), boron, boric acid, phytoestrogens, phytoandrogens, genistein, daidzein, L-carnitine, chromium pyridinecarboxate, chromium tripyridinecarboxate. Triticolinate, chromium nicotinate, acid / base modifiers, potassium citrate, potassium chloride, calcium carbonate, calcium chloride, sodium bisulfate; eucalyptus, lavender, peppermint, plasticizers, colorants, flavorings, sweeteners, buffers, lubricants, carriers, pH adjusters, natural ingredients, stabilizers, biological additives such as enzymes (including proteases and lipases), chemical additives, coolants, chelating agents, denaturants, astringents, emulsifiers, topical analgesics, fragrance compounds, humectants, emulsifiers. (e.g., zinc oxide and titanium dioxide), defoamers (e.g., silicone), preservatives (e.g., butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), propyl gallate, benzalkonium chloride, EDTA, benzyl alcohol, potassium sorbate, parabens and mixtures thereof), reducing agents, solvents, co-solubilizers, solubilizers, suspending agents (non-surfactants), solvents, thickeners (aqueous and non-aqueous), complexing agents, keratolytic agents, eggshell membranes, ancient grains and combinations thereof.

[0078] Pet food compositions can be produced by various methods to achieve a desired pet food composition or a desired form of pet food composition. For example, dry food can be baked or extruded and then cut into individual shaped portions, such as extruded food. In some embodiments, pet food compositions can be prepared in canned or wet form using conventional food preparation processes known to those skilled in the art. Typically, milled animal protein tissue is mixed with other ingredients, such as grain flour cereals, suitable carbohydrate sources, fats, oils, and balancing ingredients (including special-purpose additives such as vitamin and mineral mixtures, inorganic salts, cellulose, and beet pulp), and water in an amount sufficient for processing. The ingredients are mixed in a container suitable for heating while simultaneously blending the components. Heating of the mixture is carried out using any suitable method, such as direct steam injection or using a container equipped with a heat exchanger. After all the ingredients of the formulation have been added, the mixture can be heated to a temperature of 50℉ to 212℉. Although temperatures outside this range can be used, this may be commercially impractical without the use of other processing aids. When heated to the appropriate temperature, the material will typically be in the form of a viscous liquid, which is dispensed into the can. A lid is applied and the container is hermetically sealed. The sealed container is then placed in conventional equipment designed for sterilizing the contents. Sterilization is typically accomplished by heating to a temperature exceeding 230°C for an appropriate time, the appropriate time depending on the temperature used, the nature of the composition, and relevant factors. The pet food compositions and food products disclosed herein can also be added to or combined with food compositions before, during, or after their preparation.

[0079] In some embodiments, the pet food compositions disclosed herein may be prepared in dry form using conventional processes known to those skilled in the art. Typically, dry ingredients, including dried animal proteins, plant proteins, grains, etc., are ground and mixed together. Liquid or moist ingredients, including fats, oils, animal proteins, water, etc., are added together with the dry materials. The specific formulation, order of addition, combination, methods, and equipment used to combine the various ingredients may be selected from those known in the art. For example, in some embodiments, the resulting mixture is processed into extruded food or similar dry blocks formed using an extrusion process in which the mixture of dry and wet ingredients is subjected to mechanical work under high pressure and high temperature, forced through small openings or orifices, and cut into extruded food, for example, by a rotary cutter. The resulting extruded food may then be dried and optionally coated with one or more partial coatings, the partial coatings comprising, for example, flavorings, fats, oils, powdered ingredients, etc. Extruded food may also be prepared from dough by baking rather than extrusion, wherein the dough is placed in a mold and then subjected to dry heat treatment.

[0080] In preparing the composition, any component can typically be incorporated into the composition during the formulation process, such as during and / or after mixing with other components of the composition. The dispensing of these components into the composition can be accomplished in a conventional manner. In some embodiments, milled animal and / or poultry protein tissue is mixed with other components, including nutritional balancers, inorganic salts, and may further include cellulose, beet pulp, growth promoters, etc., along with sufficient water for treatment.

[0081] In some embodiments, the ingredients of the pet food composition disclosed herein are extruded and then coated with a surface coating comprising D-ribose and optionally palatability enhancers and / or other coating ingredients. In some embodiments, the extruded food is coated together with the composition comprising D-ribose and optionally palatability enhancers and / or other coating ingredients in a rotary mixer. In other embodiments, the extruded food is coated with a coating comprising D-ribose using a vacuum coating technique, wherein the extruded food is subjected to a vacuum and then exposed to the coating material, after which the release of the vacuum drives the coating material within the extruded food. In some embodiments, the uncoated extruded food comprises D-ribose, and the uncoated extruded food may remain uncoated or may subsequently be coated, for example, with palatability enhancers in any manner disclosed herein.

[0082] According to certain embodiments, this document further discloses a method for treating, preventing, and / or alleviating periodontal disease in companion animals, the method comprising administering to the companion animal in need a composition as disclosed herein.

[0083] According to another aspect, a method for reducing dental plaque and / or tartar in companion animals is provided, the method comprising administering to the companion animal in need a composition as disclosed herein.

[0084] According to yet another embodiment, this document discloses a method for reducing the abundance of at least one of Fusobacterium nucleatum or Porphyromonas gingivalis in the oral cavity of a companion animal, the method comprising administering to the companion animal in need a composition as disclosed herein.

[0085] Any method disclosed herein may include providing and / or feeding a companion animal (e.g., a dog) with a pet food composition as disclosed herein. In some cases, the method may include providing and / or feeding the companion animal with the pet food composition for 1 day or more, preferably 5 days or more, preferably 7 days or more, preferably 10 days or more, preferably 14 days or more, preferably 30 days or more, or preferably 42 days or more. The method may include feeding the companion animal once a day, twice a day, three times a day, or in some embodiments four or more times a day.

[0086] Alternatively or concurrently, any method disclosed herein may include applying and / or dispensing a dental cleaning agent as disclosed herein to the mouth of a companion animal (e.g., a dog). In some cases, the dental cleaning agent may be applied to the mouth of the companion animal for one or more days, preferably five or more days, preferably seven or more days, preferably ten or more days, preferably fourteen or more days, preferably thirty or more days, or preferably for forty-two or more days. The method may include applying the dental cleaning agent to the mouth of the companion animal once daily, twice daily, once weekly, twice weekly, three times weekly, four times weekly, or once monthly.

[0087] The examples and other embodiments described herein are exemplary and are not intended to limit the scope of the compositions and methods described herein. Equivalent changes, modifications, and variations can be made to the specific embodiments, materials, compositions, and methods within the scope of this disclosure to obtain substantially similar results.

[0088] Example

[0089] Example 1

[0090] Four exemplary pet food compositions were prepared according to various aspects of this disclosure as follows: (1) a negative control food; (2) a positive control food; (3) test food 1; and (4) test food 2, wherein all four foods were formulated according to AAFCO nutritional recommendations and were in the form of extruded food produced by an extrusion method. The negative control food contained neither mechanical nor chemical components to achieve the destruction of dental plaque buildup and was dried and coated with an attractant after extrusion. The positive control food was an extruded food that was dried and coated with an attractant after extrusion. Unlike the negative control food and test food 1 or test food 2, the positive control food was produced following a method with a transverse fibrous striation matrix that provided a mechanical cleaning effect (to simulate brushing), which had previously been shown to reduce dental plaque buildup. Test food 1 was dried and coated with an attractant after extrusion and formulated with prebiotic fiber bundles containing pecan shell fiber, cranberry pomace, citrus pulp, beet pulp, and flaxseed. Test food 2 was formulated to include 2% D-ribose, which was first mixed with a liquid palatability enhancer and thoroughly stirred, and then coated onto extruded and dried puffed food. Tables 1 and 2 below show four pet food compositions and their predicted approximate analyses, respectively.

[0091] Table 1 - Formulation of Pet Food Compositions

[0092]

[0093]

[0094] Table 2 - Nutritional analysis of pet food compositions (predicted by PDM)

[0095]

[0096]

[0097] This feeding study was conducted on a total of 20 clinically healthy adult dogs (based on blood CBC / serum chemistry and urinalysis), weighing between 12.59 kg and 22.66 kg, and aged between 4 years and 7 months and 7 years and 11 months at the start of the study. All dogs were mixed in terms of sex and reproductive status (unneutered, castrated, or spayed). Briefly, based on sex, age, and a full-oral plaque score measured and considered as baseline before the start of the feeding study, the 20 dogs were randomly assigned to four groups of five dogs each. Figure 1 As shown, the feeding study was conducted using William's Latin Square design, which resulted in each subject receiving each of the four food compositions over a total of 28 days.

[0098] Before initiating the feeding trial, a full oral scaling (supragingival and subgingival) and dental examination were performed. Biofilm removal was confirmed at the supragingival and subgingival levels during scaling using eosin dye or a dental flashlight.

[0099] After dental cleaning, follow the instructions. Figure 1The diets were fed to each group in the specified order, with each diet administered for 28 days. On day 28, plaque and tartar were graded under anesthesia using the Logan-Boyce method. Total plaque and tartar accumulation was quantified using Veterinary Oral Health Council (VEHC) indicators and those applied to the “whole teeth” (plaque and tartar not separated) as outlined below. Plaque on each tooth was scored using the following numerical scores based on coverage and thickness (eosin staining intensity): Coverage: 0 = No plaque detected; 1 = 1–24%; 2 = 25–49%; 3 = 50–74%; 4 = 75–100%. Thickness: 1 = Mild; pink to light red; 2 = Moderate, red. Tartar was scored for each tooth based on coverage using the following numerical scores: Coverage: 0 = No tartar detected; 1 = 1-24%; 2 = 25-49%; 3 = 50-74%; 4 = 75-100%. Gingivitis scores for each tooth were measured using the Loe & Silness method, using the following indicators: 0 = Normal gingiva; 0.5 = Mild inflammation with slight redness; 1 = Moderate inflammation and redness, but no bleeding on probing; 2 = Moderate inflammation with severe redness and bleeding on probing; and 3 = Severe inflammation and redness, edema, ulceration, and spontaneous bleeding. Average scores for plaque, tartar, and gingivitis were calculated by averaging the total tooth scores for each animal. After tooth grading, all dogs underwent full-mouth dental cleaning, followed by a different diet for an additional 28 days, and so on, to complete the following procedures. Figure 1 The order is shown. The four diets described in Tables 1 and 2 above were used in this study. All dogs were allowed regular activities, except that chew toys or treats were not permitted.

[0100] Subgingival plaque sample collection: Subgingival plaque samples were collected for microbiome analysis. For each subject, subgingival plaque samples were collected from both healthy and unhealthy teeth of each dog. Healthy and unhealthy teeth were identified based on individual gingivitis scores. A gingivitis score of "0 = normal gingiva" was considered a healthy tooth, and a gingivitis score of 0.5 or higher was considered an unhealthy tooth. If the subject had no unhealthy teeth, subgingival plaque samples were collected from two different healthy teeth. Similarly, if the subject had no healthy teeth, subgingival plaque samples were collected from two different unhealthy teeth.

[0101] Microbiome sequencing analysis of subgingival plaque samples was performed using total DNA extracted from frozen samples using the Qiagen DNeasy Power Biofilm Extraction Kit. Following total DNA extraction, 16S rDNA amplicones were prepared from the samples using a primer set spanning the V1 and V3 hypervariable regions via PCR, and subsequently qualitatively analyzed using an Agilent 2100 Bioanalyzer. Indexing PCR was also performed, followed by library quantification, normalization, and pooling of the samples, with modifications following the manufacturer's instructions. The final pooled sample library was loaded into the MiSeq v3 Sample Loading Kit, and the kit was placed in an Illumina MiSeq sequencer for amplicon sequencing. The sample sequences were then split using MiSeq's built-in metagenomics workflow to obtain FASTQ files. The FASTQ files were processed using Mothur software to classify the sequence reads using the Human Oral Microbiome Database (HOMD v.15.11), followed by custom modifications.

[0102] Data filtering and statistical analysis: For the microbiome analysis, abundance data were filtered by prevalence (OTUs representing >25% of samples in the dataset) and read coverage (>98.0% of sequencing reads were retained per sample; 83,558.86 ± 42,799.45 reads per sample). After filtering, count data were transformed to median log-ratio (CLR) values ​​for statistical analysis applicable to the composition data. The study was a crossover design, and therefore a mixed model analysis was performed using JMP Pro v.17.0 with food type as a fixed effect and animal identification as a random effect to handle repeated measures data with false discovery rate (FDR) corrected p-values. Tucky post-hoc tests were used to assess paired differences between diet types for each variable.

[0103] Results: Based on the mixed model analysis, the "food effect" showed a significant impact on overall dental plaque scores (p = < 0.0001), but the "food feeding order effect" was not observed. Tuki post-hoc significance tests showed that baseline overall dental plaque scores were higher compared to other foods. For overall dental plaque scores, dogs fed the positive control food showed the largest significant reduction, followed by test food 2, then test food 1, and baseline, as shown below. Figure 2As shown. Without being bound by theory, it is believed that the greatest reduction in overall plaque score was observed in the positive control food, due to the mechanical effect provided by the puffed food, which was prepared using a specialized food processing technique (as described above). Similarly, these findings support the conclusion that the significant reduction in overall plaque score demonstrated by test food 2 is due to the chemical effect provided by including D-ribose, which is added during the coating stage of the finished puffed food. Overall, test food 2 showed a significant reduction in overall plaque score compared to the negative control food and test food 1.

[0104] In addition to reducing overall dental plaque scores, dogs fed test food 2 also showed a non-significant decreasing trend line in the abundance levels of specific microbial species (including *Fusobacterium nucleatum* subsp. *vincentii* and *Porphyromonas gingivalis*) in subgingival plaque compared to dogs fed other food compositions, as shown in Figures 1 and 2. Figure 4 As shown, *Fusobacterium nucleatum*, a member of the "orange complex," is a common member of the oral microbiota in both periodontal health and disease, and acts as a bridging species to metabolically connect early and late bacterial colonizing bacteria (i.e., potential pathogen "red complex" species) during plaque maturation. Furthermore, the abundance level of *Fusobacterium nucleatum* was significantly higher in unhealthy teeth (gingivitis score > 0.5) compared to healthy teeth (gingivitis score < 0.05). Figure 3 As shown.

[0105] Fusobacterium nucleatum also plays a role in halitosis, pulpitis, and oral cancer. Furthermore, Fusobacterium nucleatum promotes the attachment of the "red complex" bacterial species *Porphyromonas gingivalis* by activating NADPH oxidase isoforms (NOX1 and NOX2) in human gingival fibroblasts. It is believed that the formation of this microbial complex through these two species contributes to the pathogenesis of periodontal disease. Interestingly, compared to dogs fed other diets, dogs fed test diet 2 showed a non-significant decreasing trend in the abundance of *Porphyromonas gingivalis* in subgingival plaque, as shown in the figure. Figure 4 As shown, the abundance level of Porphyromonas gingivalis in healthy teeth was also significantly reduced compared to unhealthy teeth. Figure 4 Compared to healthy teeth (gingivitis score < 0.5), unhealthy teeth (based on a gingivitis score of 0.5) showed significantly higher abundance levels of both *Fusobacterium nucleatum* and *Porphyromonas gingivalis*. However, a considerable number of teeth showed higher plaque scores, ranging from 4 to 8. Figure 5As shown, this indicates that the development of gingivitis and progression to periodontitis is more related to the presence of key microbial species than to plaque buildup itself. *Porphyromonas gingivalis* is considered a key pathogen, disrupting host symbiotic homeostasis and ultimately interfering with the host's innate immunity to periodontitis. The prevalence of *Porphyromonas gingivalis* is higher in dogs (88%) and cats (96%) than other potential periodontal pathogens such as *Porphyromonas gulae* (39% in dogs, 70% in cats) and *Tannerella forsythia* (4% in dogs, 2% in cats).

[0106] like Figure 6 As shown, *Fusobacterium nucleatum* showed a significant positive correlation with gingivitis scores (R² = 0.206, P < 0.001), but no significant relationship was found with plaque or tartar scores. Similarly, *Fusobacterium nucleatum* showed a non-significant positive correlation with overall gingivitis scores, but a non-significant negative correlation with overall plaque and tartar scores, such as... Figure 7 As shown. However, *Porphyromonas gingivalis* showed a significant positive correlation with gingivitis scores (R-squared = 0.167, p < 0.001), but did not show a positive correlation with dental plaque and tartar scores, as... Figure 8 As shown. Similarly, as Figure 9 As shown, *Porphyromonas gingivalis* showed a non-significant positive correlation with scores for plaque, tartar, and gingivitis throughout the mouth.

[0107] The aforementioned findings support the conclusion that, compared to a comparative food containing no D-ribose, the test food 2, which contains D-ribose as an ingredient, improved oral health by significantly reducing overall dental plaque scores and modulating subgingival microbiome composition by specifically reducing the abundance levels of key microbial species Fusobacterium nucleatum and Porphyromonas gingivalis.

Claims

1. A method for treating, preventing, or alleviating periodontal disease in companion animals, the method comprising administering to the companion animal in need a composition comprising an effective amount of D-ribose.

2. The method of claim 1, wherein the D-ribose is present in the composition in an amount ranging from about 1 wt% to about 5 wt%, preferably from about 2 wt% to about 4 wt% or about 2 wt%, based on the total weight of the composition.

3. The method according to claim 1 or 2, wherein the periodontal disease of the companion animal is treated or alleviated by reducing the amount of at least one of tartar or plaque on the teeth of the companion animal compared to before the composition is applied to the companion animal.

4. The method according to any one of the preceding claims, wherein the method reduces at least one of Fusobacterium nucleatum or Porphyromonas gingivalis present on the teeth of the companion animal compared to before the composition is applied to the companion animal.

5. The method according to any one of the preceding claims, wherein the composition is a pet food composition selected from the following forms: dry extruded food, wet cakes, dental chews, or treats.

6. The method of claim 5, wherein the pet food composition is in the form of dry extruded food.

7. The method of claim 6, wherein the D-ribose is in the form of a coating covering the dry puffed grain.

8. The method according to any one of the preceding claims, wherein the composition further comprises at least one mechanical component for cleaning teeth.

9. The method of claim 8, wherein the at least one mechanical component for cleaning teeth comprises a transverse fibrous striation matrix.

10. The method according to any one of the preceding claims, wherein the companion animal is a dog.

11. The method according to any one of the preceding claims, wherein the composition is applied for 1 day or more, preferably 5 days or more, preferably 7 days or more, preferably 10 days or more, preferably 14 days or more, preferably 30 days or more, or preferably 42 days or more.

12. A pet food composition for treating, preventing, or alleviating periodontal disease in companion animals, the pet food composition comprising an effective amount of D-ribose, wherein the pet food composition is in the form of extruded food containing a coating comprising the D-ribose.

13. The pet food composition of claim 12, wherein the D-ribose is present in the pet food composition in an amount ranging from about 1 wt% to about 5 wt%, preferably from about 2 wt% to about 4 wt% or about 2 wt%, based on the total weight of the pet food composition.

14. The pet food composition of claim 12 or 13, wherein the pet food composition does not contain any additional sugar.

15. The pet food composition according to any one of claims 12 to 14, wherein the coating further comprises at least one palatability enhancer.

16. A method for reducing tartar buildup, the method comprising administering to a companion animal a pet food composition according to any one of claims 12 to 15.

17. A method for reducing dental plaque accumulation, the method comprising administering to a companion animal a pet food composition according to any one of claims 12 to 15.

18. A method for reducing the amount of Fusobacterium nucleatum and / or Porphyromonas gingivalis in the oral cavity, the method comprising administering to a companion animal a pet food composition according to any one of claims 12 to 15.

19. A dental cleaning composition for treating, preventing or alleviating periodontal disease in companion animals, said dental cleaning composition comprising an effective amount of D-ribose.

20. The dental cleaning composition of claim 19, wherein the D-ribose is present in the dental cleaning composition in an amount ranging from about 1 wt% to about 5 wt%, preferably from about 2 wt% to about 4 wt% or about 2 wt%, based on the total weight of the dental cleaning composition.

21. The dental cleaning composition according to claim 19 or 20, wherein the dental cleaning composition does not contain any additional sugar.

22. A method for preparing a pet food composition for treating, preventing, or alleviating symptoms of periodontal disease, the method comprising: (a) Mix the wet ingredients with the dry ingredients to form a dough; (b) Extruding the dough to form extruded puffed grains; (c) Drying the extruded puffed grain; and (d) Coating dried extruded grains with localized liquid and / or dry components. In step (a) and / or step (d), D-ribose is applied to the extruded food in an amount that effectively treats, prevents or alleviates the symptoms of periodontal disease in companion animals consuming the pet food composition.

23. The method of claim 22, wherein the D-ribose is applied to the puffed grain in step (d).

24. The method of claim 22 or 23, wherein the D-ribose is applied in an amount ranging from about 1 wt% to about 5 wt%, preferably from about 2 wt% to about 4 wt% or about 2 wt%, based on the total weight of the pet food composition.

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