Biomarker-based methods and systems for identifying companion animals at risk of digestive problems

Abstract

This specification describes methods for identifying companion animals at higher risk of developing gastrointestinal (GI) diseases / conditions. In particular, a comprehensive approach including the use of intestinal metabolites (SCFAs), physiological status (pH), and fecal scores (1-5) for identifying companion animals at higher risk of developing GI conditions / diseases is described herein. Compositions, methods, and kits for treating such identified companion animals are also described.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 493,128, filed on March 30, 2023, titled Biomarker - Based Method and System for Identifying Companion Animals Having Risk for Gastrointestinal Problems, which is hereby incorporated by reference as if fully set forth herein.

Background Art

[0002] In particular, the accurate classification of individual dogs with or without risk of gastrointestinal (GI) problems, using non - invasive sampling (e.g., fecal sampling), into "at - risk for GI" versus "not at - risk for GI" remains a significant clinical challenge.

[0003] The gut microbiota performs a number of important biochemical functions by forming a mutually beneficial symbiotic association with the host. Microbiome disruption causes dysbiosis, which has been clearly associated with many non - communicable diseases and conditions (NCDs). For example, dysbiosis is clearly associated with chronic kidney disease, cardiovascular disease, circulatory disease, inflammatory bowel disease (IBD), chronic intestinal disease, chronic kidney disease (CKD), autism, celiac disease, Crohn's disease, ulcerative colitis, prostate cancer, colorectal cancer, obesity, type 1 diabetes and type 2 diabetes, as well as some skin diseases. Dysbiosis can also affect the gut - brain axis, gut - brain - skin axis, as well as drug metabolism and toxicity therefrom. Dysbiosis also affects the host immune response.

[0004] The most influential studies have demonstrated that "dysbiosis" is the condition that causes the majority of the host pathological states described above. Recent evidence supports the development of a "dysbiosis index" based on the abundance of specific microbial species in the gut to predict host risk status for certain diseases in humans and dogs (e.g., IBD and chronic bowel disease). In addition to the microbiome, gut metabolism and physiological functions mediated by the microbiome are also important factors in the progression / maintenance and development of certain disease states. Evidence clearly demonstrates that certain metabolites, particularly short-chain fatty acids (SCFAs) and pH, are involved in the development of colorectal cancer in conjunction with the microbiome, and suggests that SCFAs and pH contribute to the state of "dysbiosis."

[0005] Three key factors—(1) the microbiome, (2) gut metabolism, and (3) physiological function—are important for the progression of GI maintenance against dysbiosis and the development of specific pathological conditions. However, the dysbiosis index developed in these studies has limited applicability because it depends entirely on the abundance of specific bacterial species.

[0006] Therefore, there is a need to develop a model that identifies whether an individual dog is at risk of developing GI problems or not, for a more comprehensive approach that takes into account multiple parameters including the microbiome, gut metabolites (e.g., SCFA), physiological state (pH), and fecal score (1-5, where 1 = liquid / watery and 5 = hard and cylindrical). Generally, a fecal score of 4-5 is desirable. [Overview of the Initiative]

[0007] This summary is intended to provide a simplified summary of some aspects of one or more embodiments of the present disclosure. Further scope to which the present disclosure is applicable will become apparent from the “Modes for Carrying Out the Invention” provided below in this specification. This summary is not an extensive overview, nor is it intended to identify key or important elements of the teachings, nor to describe the scope of the present disclosure. Rather, its purpose is merely to present one or more concepts in a simplified form as a prelude to the following “Modes for Carrying Out the Invention.”

[0008] This invention relates to identifying dogs that are at risk of GI problems or not.

[0009] In one embodiment, the disclosure relates to a method for identifying whether a subject dog is at risk of or not at risk of gastrointestinal (GI) disease. The method includes (a) analyzing a fecal sample from the subject dog to determine the values ​​of the fecal sample for one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. The method also includes comparing the values ​​obtained from (a) to control values ​​of one or more dogs without risk of GI disease for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture, (vii) fecal score, and (viii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. A subject dog is considered to be at risk of GI disease if one or more of the values ​​in the fecal sample are similar to or identical to the control values. In some embodiments, a subject dog is considered to be at risk of GI disease if each of the values ​​in the fecal sample is similar to or identical to the control values. In some embodiments, a subject dog is at risk of GI disease if one or more of the values ​​in the fecal sample are different from the control values. In some embodiments, the dogs in question are at risk of GI disease if each of the values ​​in their fecal samples differs from the control value.

[0010] In one embodiment, the disclosure relates to a kit for identifying whether a subject dog is at risk of or not of gastrointestinal (GI) disease. The kit includes at least one of the following: a reagent or apparatus for determining the pH of a fecal sample; one or more reagents or apparatus for measuring volatile carboxylic acids in a fecal sample; and one or more reagents or apparatus for determining the abundance of one or more bacteria, including Prevotellaseae, Ruminococcusceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes, in a fecal sample.

[0011] In one embodiment, the disclosure relates to a method for preventing or treating gastrointestinal (GI) disease, comprising: identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease; and, if the identifying step is concluded to be that the subject dog is at risk, administering a GI disease relief diet to the subject dog. In some embodiments, the step of identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease involves (a) analyzing a fecal sample from the subject dog to determine the values ​​of the fecal sample for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture, (vii) fecal score, and (viii) the abundance of one or more of the bacteria Prevotella seaae, Ruminococcus seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes; and (b) using the values ​​obtained from (a) to determine, respectively, (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, and (iv) C4 This includes comparing the SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viii) the abundance of one or more bacteria, including Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes, to control values ​​from one or more dogs without risk of GI disease. A dog is considered to be at no risk of GI disease if one or more values ​​in its fecal sample are similar to or identical to the control values, or to be at risk of GI disease if one or more values ​​in its fecal sample differ from the control values. In some embodiments, the GI disease mitigation diet comprises a dog food composition of quinoa, amaranth, eggshell membrane, and dietary fiber. In some embodiments, the dog food composition comprises about 1% to about 10% by weight of quinoa, about 1% to about 10% by weight of amaranth, about 0.5% to about 8% by weight of eggshell membrane, and about 2% to about 20% by weight of dietary fiber. In some embodiments, the dietary fiber includes insoluble fiber and soluble fiber. In some embodiments, the dietary fiber includes about 1% to about 18% by weight of insoluble fiber and about 0.5% to about 8% by weight of soluble fiber.The weight percentage is based on the total weight of the dog food composition on a dry matter basis.

[0012] In one embodiment, the disclosure relates to a device comprising an input, memory, a processor, and an output. The input is configured to receive the results of a method for identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease. The memory includes processor-executable instructions for comparing the results with a control value, for preparing a conclusion that the dog is at risk or not based on the comparison, and for outputting the results. The processor is configured to execute the processor-executable instructions. The output is configured to display the conclusion. In some embodiments, the processor-executable instructions further include instructions that display a recommendation to administer a GI disease relief diet to the subject when the subject dog is concluded to be at risk, the processor is further configured to execute the instructions and display the recommendation, and the output is further configured to display the recommendation.

[0013] In one embodiment, a method is provided for identifying whether a subject dog is at risk of or not of gastrointestinal (GI) disease, comprising (a) analyzing a fecal sample from the dog for three or more of the following: pH, total short-chain fatty acid content (SCFA), and C2 SCFA, C3 SFFA, C4 SCFA, C5 SCFA, and / or C6 SCFA content, fecal score, moisture content, and the relative abundance of phylum bacteroidetes excluding Prevotella seaae, F3:f_bacteroideda seaae and F4:f_prevotella seaae, Ruminococca seaae, Enterobacteria seaae, Clostridiares and Prevotella, and any other bacterial phylum, family, genus or species. The method also comprises (b) comparing the values ​​obtained from (a) with a control value obtained from one dog or a pooled control value from multiple dogs that have been medically determined not to be at risk of GI disease. A dog is identified as being at no risk of GI disease if its fecal sample has the following characteristics: (i) pH at or below the control level, and total SFCA at or above the control level; and (ii) moisture and / or Enterobacteria seaae at or below the control level when measured, and levels of C3 SCFA, C2 SFCA, C4 SCFA, fecal score, Prevotella seaae, Ruminococca seaae, Clostridiares and / or Prevotella at or above the control level. The presence of other bacteria may also be determined, as greater diversity in the gut microbiome is also generally associated with dogs at no risk.

[0014] In one embodiment, a kit is provided for identifying whether a subject dog is at risk of or not of gastrointestinal (GI) disease. The kit comprises a reagent for determining the pH of a fecal sample, one or more reagents for measuring volatile carboxylic acids in the fecal sample, and one or more reagents for quantitatively identifying one or more of the following in the fecal sample: Prevotella seaae, F3:f_bacteroidaceae and F4:f_prevotella seaae, Filam bacteroidetes, luminococcus seaae, Enterobacteria seaae, Clostridiales, and / or Prevotella. In some embodiments, the method further comprises quantitatively identifying other bacteria.

[0015] In one embodiment, as further described herein, a method is provided for preventing or treating GI disease, comprising identifying the GI status of a dog and administering a GI disease mitigation diet to dogs identified as being at risk of GI disease. [Brief explanation of the drawing]

[0016] The features and advantages of the present invention will become apparent from the following more detailed "Modes for Carrying Out the Invention" concerning specific embodiments of the present invention, and as illustrated in the accompanying drawings.

[0017] [Figure 1A-1B] Figures 1A and 1B show the principal component analysis of 96 variables derived from pH, SCFA, microbiome, and other metadata.

[0018] [Figure 2] Figure 2 shows the receiver operational characteristics (ROC) of the discovery cohort and associated confusion matrix for prediction.

[0019] [Figure 3A-3B]Figures 3A and 3B show the abundance of important variables used in a model that distinguishes individuals into two groups (with GI risk vs. without GI risk). Microbiome variables were represented as relative abundances (centered log ratio transformation), and other variables were represented by their actual levels. SCFAs were represented in ppm, and moisture was represented in %.

[0020] [Figure 4A-4B] Figures 4A and 4B show principal component analysis (PCA) using selected discriminators to distinguish between without GI risk (white diamonds) and with GI risk (black circles).

[0021] [Figure 5A-5B] Figures 5A and 5B show scatter plots of important variables that have a significant correlation with pH. White diamonds = without GI risk, black circles = with GI risk. Microbiome variables were represented as relative abundances, and other variables were represented by their actual levels. **DETAILED DESCRIPTION OF THE INVENTION**

[0022] For purposes of illustration, the principles of the present invention are described by reference to its various exemplary embodiments. Although specific embodiments of the present invention are specifically described herein, those skilled in the art will readily recognize that the same principles are equally applicable and can be employed in other compositions and methods. Before explaining the disclosed embodiments of the present invention in detail, it is understood that the present invention is not necessarily limited to the details of any particular disclosed embodiment in its application. The terms used herein are for purposes of description and not for purposes of limitation.

[0023] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include multiple references unless otherwise specified by context. The singular form of any class of components refers not only to one component in that class but also to a mixture of those components. The terms “a” (or “an”), “one or more,” and “at least one” may be used interchangeably herein. The terms “comprising,” “including,” and “having” may be used interchangeably. A list referred to as “at least one of A, B, and C” or “at least one of A, B, or C” means one or more of A, B, and C, not at least one A, at least one B, and / or at least one C. The term “include” should be interpreted as “includes but not limited to these.” The term “including” should be interpreted as “includes but not limited to these” or “includes but not limited to these.”

[0024] As used throughout, the term "range" is used as a shorthand to describe each and all values ​​within that range. Any value within the range can be selected as the upper and lower limits of that range. Thus, the range 1–5 specifically includes 1, 2, 3, 4, and 5, as well as subranges including 1–4, 1–3, 1–2, 2–5, 2–4, 2–3, 3–5, 3–4, and 4–5. The term "about" when referring to a number means any number within a 15% range of that number. Further embodiments of this specification include removing the word "about" in one or more cases in embodiments of this specification that include "about". Further embodiments of this specification include removing the word "about" in all cases in embodiments of this specification that include "about".

[0025] Abbreviations and symbols used in this specification have their ordinary meanings unless otherwise indicated. The abbreviation "wt.%" means weight percent relative to the pet food composition. The symbol "°" refers to degrees, such as degrees of temperature or degrees of an angle. The symbols "h", "min", "mL", "nm", "μm" mean hour, minute, milliliter, nanometer, and micrometer, respectively. The abbreviation "UV-VIS" referring to a spectrometer or spectroscopy means ultraviolet-visible. The abbreviation "rpm" means revolutions per minute.

[0026] All references cited within this specification are hereby incorporated by reference in their entirety. In case of any conflict between the definitions in the present disclosure and those in the cited references, the present disclosure prevails.

[0027] One embodiment includes a method for identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease. The method includes (a) analyzing a fecal sample from the subject dog to determine the values ​​of the fecal sample for one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. The method also includes comparing the values ​​obtained from (a) to control values ​​of one or more dogs without risk of GI disease for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture, (vii) fecal score, and (viii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. The control values ​​may be obtained from a reference source or table of previously obtained control values. The control values ​​may be from this specification. Control values ​​can be obtained from experiments involving one or more dogs without risk of GI disease, which are performed using the same methods as those used to determine the values ​​of fecal samples from control dogs for one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA levels, (iii) C3 SCFA levels, (iv) C4 SCFA levels, (v) pH, (vi) moisture content, (vii) fecal score, and (viii) the abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes, but with the analysis of fecal samples from control dogs(s).

[0028] In some embodiments, a dog is determined to be at no risk of GI disease if one or more values ​​in its fecal sample are similar to or identical to the control value.

[0029] In some embodiments, step (a) of the analysis is performed to determine the values ​​of the fecal sample for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. In some embodiments, (b) comparison is made with a control value for one of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture, (vii) fecal score, and (viii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes.

[0030] In some embodiments, a dog is determined to be at no risk of GI disease if each of the fecal sample values ​​is similar to or identical to the control value. In some embodiments, a dog is determined to be at risk of GI disease if each of the fecal sample values ​​is different from the control value.

[0031] In some embodiments, a dog is determined to be at no risk of GI disease if its fecal sample has, compared to a control value, (i) a total short-chain fatty acid content at approximately the same or higher level, (ii) a total C2 SCFA at approximately the same or higher level, (iii) a total C3 SCFA at approximately the same or higher level, (iv) a total C4 SCFA at approximately the same or higher level, (v) a pH at approximately the same or lower level, (vi) a total moisture content at approximately the same or lower level, (vii) a total fecal score at approximately the same or higher level, and (viii) a total abundance of Prevotellaceae, Ruminococcaceae, and Bacteroidaceae bacteria at approximately the same or higher level, and a total abundance of Enterobacteriaceae and Bacteroidetes bacteria other than Prevotellaceae and Bacteroidaceae at approximately the same or lower level.

[0032] In some embodiments, a dog is determined to be at risk of GI disease if its fecal sample has, compared to a control value, (i) a lower total content of short-chain fatty acids, (ii) a lower level of C2 SCFA, (iii) a lower level of C3 SCFA, (iv) a lower level of C4 SCFA, (v) a higher pH, (vi) more moisture, (vii) a lower fecal score, and (viii) a lower abundance of Prevotellaceae, Ruminococcaceae, and Bacteroidaceae bacteria, and approximately a higher abundance of Enterobacteriaceae and Bacteroidetes bacteria other than Prevotellaceae and Bacteroidaceae.

[0033] In some embodiments, a dog is determined to be at no risk of GI disease if, compared to a control value, its fecal sample has: (i) a total short-chain fatty acid content at approximately the same or higher level; (ii) a total short-chain fatty acid content at approximately the same or higher level; (iii) a total short-chain fatty acid content at approximately the same or higher level; (iii) a total short-chain fatty acid content at approximately the same or higher level; (iv) a total short-chain fatty acid content at approximately the same or lower level; (v) a total short-chain fatty acid content at approximately the same or lower level; (vi) a total short-chain fatty acid content at approximately the same or lower level; (vii) a total short-chain fatty acid content at approximately the same or higher

[0034] In some embodiments, a dog is determined to be at risk of GI disease if its fecal sample has, compared to a control value, (i) a lower total content of short-chain fatty acids, (ii) a lower level of C2 SCFA, (iii) a lower level of C3 SCFA, (iv) a lower level of C4 SCFA, (v) a higher pH, (vi) more moisture, (vii) a lower fecal score, or (viii) a lower abundance of Prevotellaceae, Ruminococcaceae, and Bacteroidaceae bacteria, and at least one, two, three, four, five, six, or seven of the Enterobacteriaceae and Bacteroidetes bacteria, one or both of which are approximately higher than Prevotellaceae and Bacteroidaceae.

[0035] One embodiment includes a method for identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease. The method includes (a) analyzing a fecal sample from the subject dog to determine the values ​​of the fecal sample for one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. A dog is considered to be free from the risk of GI disease if the values ​​of its fecal sample include (i) the total content of short-chain fatty acids (SCFA) found in dogs without risk in the examples herein, (ii) the level of C2 SCFA found in dogs without risk in the examples herein, (iii) the level of C3 SCFA found in dogs without risk in the examples herein, (iv) the level of C4 SCFA found in dogs without risk in the examples herein, (v) the pH found in dogs without risk in the examples herein, (vi) the moisture content found in dogs without risk in the examples herein, (vii) the fecal score found in dogs without risk in the examples herein, and (vii) the abundance of one or more bacteria from among Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes found in dogs without risk in the examples herein. In some embodiments, the values ​​of the fecal sample include each of (i) to (vii). In some embodiments, the values ​​of the fecal sample include each of (i) to (vii), where the value of (viii) includes the abundance of each of the bacteria Prevotellaceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes.

[0036] In some embodiments, a dog is determined to be at risk of GI disease if its fecal sample contains one or more of the following values: (i) a total short-chain fatty acid content (SCFA) lower than that observed in dogs without risk in the examples herein; (ii) a lower level of C2 SCFA than that observed in dogs without risk in the examples herein; (iii) a lower level of C3 SCFA than that observed in dogs without risk in the examples herein; and (iv) a lower level of C4 SCFA than that observed in dogs without risk in the examples herein. SCFA, (v) pH higher than that observed in dogs without risk in the examples herein, (vi) moisture higher than that observed in dogs without risk in the examples herein, (vii) fecal score lower than that observed in dogs without risk in the examples herein, and (viii) one or more of the following: a lower abundance of one or more bacteria from Prevotellaceae, Ruminococcaceae, and Bacteroidetes, and a higher abundance of one or both of Enterobacteriaceae and Bacteroidetes other than Prevotellaceae and Bacteroidetes, than that observed in dogs without risk in the examples herein. In some embodiments, the values ​​of the fecal sample include each of (i) to (vii). In some embodiments, the fecal sample values ​​include each of (i) to (vii), where value (viii) includes the respective abundances of Prevotella seaae, Ruminococca seaae, and Bacteroida seaae bacteria, which are lower than those observed in risk-free dogs in the examples herein, and the respective abundances of Enterobacteria seaae and Bacteroidetes bacteria, which are higher than those observed in risk-free dogs in the examples herein.

[0037] In some embodiments, the method for determining any of (i) to (vii), as well as any one of the bacteria Prevotellaceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes, may be as described in the examples herein.

[0038] One embodiment includes a kit for identifying whether a subject dog is at risk of or not of gastrointestinal (GI) disease by obtaining the results of a fecal analysis. The kit includes a reagent or apparatus for determining the pH of a fecal sample, one or more reagents or apparatus for measuring volatile carboxylic acids in the fecal sample, and at least one of one or more reagents or apparatus for determining the abundance of one or more bacteria from among Prevotellaseae, Luminococcusceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes in the fecal sample. The results of the fecal analysis are obtained by implementing the aforementioned components of the kit.

[0039] In some embodiments, the kit includes each of the reagents or devices for determining the pH of the fecal sample, one or more reagents or devices for measuring volatile carboxylic acids in the fecal sample, and one or more reagents or devices for determining the abundance of each of the following bacteria in the fecal sample: Bacteroidetes, Prevotellaceae, Ruminococcaceae, Enterobacteriaceae, Bacteroidaceae, and Prevotella.

[0040] In some embodiments, the kit includes one or more reagents for determining the abundance of Enterobacteria seaae and Prevotella seaae in a sample.

[0041] In some embodiments, one or more reagents or apparatus for determining the abundance of one or more bacteria from Bacteroidetes, Prevotellaceae, Ruminococcaceae, Enterobacteriaceae, Bacteroidaceae, and Prevotella in a fecal sample includes PCR primers for amplifying 16S rDNA from the target bacteria.

[0042] In some embodiments, the kit further includes instructions for analyzing the results of a fecal sample analysis of a dog under consideration using the kit. In some embodiments, the instructions for analysis include concluding that the dog under consideration is at risk of gastrointestinal (GI) disease if the values ​​of the fecal sample include one or more of the following: (i) a total short-chain fatty acid content (SCFA) lower than that observed in dogs without risk in the examples herein; (ii) a lower level of C2 SCFA than that observed in dogs without risk in the examples herein; (iii) a lower level of C3 SCFA than that observed in dogs without risk in the examples herein; (iv) a lower level of C4 SCFA than that observed in dogs without risk in the examples herein. SCFA, (v) pH higher than that observed in dogs without risk in the examples herein, (vi) moisture higher than that observed in dogs without risk in the examples herein, (vii) fecal score lower than that observed in dogs without risk in the examples herein, and (viii) one or more of the following: a lower abundance of one or more bacteria from Prevotellaceae, Ruminococcaceae, and Bacteroidaceae than that observed in dogs without risk in the examples herein, and a higher abundance of one or both of Enterobacteriaceae and Bacteroidetes other than Prevotellaceae and Bacteroidaceae than that observed in dogs without risk in the examples herein.In some embodiments, the instructions for analysis include concluding that a dog is at risk of gastrointestinal (GI) disease if the values ​​of the fecal sample include any of the following: (i) a total short-chain fatty acid content (SCFA) lower than that observed in dogs without risk in the examples herein; (ii) a lower level of C2 SCFA than that observed in dogs without risk in the examples herein; (iii) a lower level of C3 SCFA than that observed in dogs without risk in the examples herein; and (iv) a lower level of C4 SCFA than that observed in dogs without risk in the examples herein. SCFA, (v) pH higher than that observed in dogs without risk in the examples herein, (vi) moisture higher than that observed in dogs without risk in the examples herein, (vii) fecal score lower than that observed in dogs without risk in the examples herein, and (viii) one or more of the following: a lower abundance of one or more bacteria from Prevotellaceae, Ruminococcaceae, and Bacteroidaceae than that observed in dogs without risk in the examples herein, and a higher abundance of one or both of Enterobacteriaceae and Bacteroidetes other than Prevotellaceae and Bacteroidaceae than that observed in dogs without risk in the examples herein.In some embodiments, the instructions for analysis include concluding that the dog is not at risk of gastrointestinal (GI) disease if the values ​​of the fecal sample include (i) the total content of short-chain fatty acids (SCFA) found in dogs without risk in the examples herein, (ii) the level of C2 SCFA found in dogs without risk in the examples herein, (iii) the level of C3 SCFA found in dogs without risk in the examples herein, (iv) the level of C4 SCFA found in dogs without risk in the examples herein, (v) the pH found in dogs without risk in the examples herein, (vi) the moisture content found in dogs without risk in the examples herein, (vii) the fecal score found in dogs without risk in the examples herein, and (vii) the abundance of one or more bacteria from among Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes found in dogs without risk in the examples herein. In some embodiments, the instructions for analysis include concluding that the dog is not at risk of gastrointestinal (GI) disease if the values ​​of the fecal sample include (i) the total content of short-chain fatty acids (SCFA) found in dogs without risk in the examples herein, (ii) the level of C2 SCFA found in dogs without risk in the examples herein, (iii) the level of C3 SCFA found in dogs without risk in the examples herein, (iv) the level of C4 SCFA found in dogs without risk in the examples herein, (v) the pH found in dogs without risk in the examples herein, (vi) the moisture content found in dogs without risk in the examples herein, (vii) the fecal score found in dogs without risk in the examples herein, and (vii) the abundance of one or more bacteria from among Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes found in dogs without risk in the examples herein.

[0043] In some embodiments, the instructions outline a method for identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease by performing the elements of the kit. The method includes (a) analyzing a fecal sample from the subject dog to determine the values ​​of the fecal sample for (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (vii) the abundance of one or more of the bacteria Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. The method also includes comparing the values ​​obtained from (a) to control values ​​of one or more dogs without risk of GI disease for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture, (vii) fecal score, and (vii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. The control values ​​may be obtained from a reference source or table of previously obtained control values. The control values ​​may be from this specification. Control values ​​can be obtained from experiments involving one or more dogs without risk of GI disease, which are performed using the same methods as those used to analyze fecal samples from control dogs to determine the values ​​of one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA levels, (iii) C3 SCFA levels, (iv) C4 SCFA levels, (v) pH, (vi) moisture content, (vii) fecal score, and (vii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes, but with the analysis of fecal samples from control dogs(s).

[0044] In some embodiments, the instructions outline a method for identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease by performing the elements of the kit. The method includes (a) analyzing a fecal sample from the subject dog to determine the values ​​of the fecal sample for (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (vii) the abundance of one or more of the bacteria Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. A dog is considered to be free from the risk of GI disease if the values ​​of its fecal sample include (i) the total content of short-chain fatty acids (SCFA) found in dogs without risk in the examples herein, (ii) the level of C2 SCFA found in dogs without risk in the examples herein, (iii) the level of C3 SCFA found in dogs without risk in the examples herein, (iv) the level of C4 SCFA found in dogs without risk in the examples herein, (v) the pH found in dogs without risk in the examples herein, (vi) the moisture content found in dogs without risk in the examples herein, (vii) the fecal score found in dogs without risk in the examples herein, and (viii) the abundance of one or more bacteria from among Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes found in dogs without risk in the examples herein. In some embodiments, the values ​​of the fecal sample include each of (i) to (viii). In some embodiments, the values ​​of the fecal sample include each of (i) to (vii), where the value of (viii) includes each of the abundances of Prevotellaceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes found in dogs without risk in the examples herein.

[0045] In some embodiments, the kit further comprises an apparatus including an input, memory, a processor, and an output. The input is configured to receive the results of any method described herein. In some embodiments, the input is configured to receive the results of a method performed while implementing other components of the kit. The memory includes processor-executable instructions for comparing the results with a control value and preparing a conclusion that there is or is not risk based on the comparison, and for outputting the results. The processor is configured to execute the processor-executable instructions. The output is configured to display the conclusion. In some embodiments, the processor-executable instructions further include instructions that, if the conclusion is that the subject dog is at risk, display a recommendation to administer a GI disease relief diet to the subject, and the output is further configured to display the recommendation.

[0046] One embodiment includes a method for preventing or treating GI disease. The method includes identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease. The method also includes administering a GI disease relief diet to the subject dog when the identifying step concludes that the subject dog is at risk. The method for identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease may be as described herein.

[0047] In some embodiments, in a method for preventing or treating GI disease, the step of identifying a subject dog as being at risk or not at risk of gastrointestinal (GI) disease includes one or more of the following: (a) analyzing a fecal sample from the subject dog to determine the values ​​of the fecal sample for one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viii) the abundance of bacteria Prevotella seaae, Ruminococcus seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. In some embodiments, the method also includes comparing the values ​​obtained from (b)(a) to control values ​​of one or more dogs without risk of GI disease for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. In some embodiments, a dog is determined to be at risk of GI disease if one or more values ​​in the fecal sample are similar to or equivalent to the control values. In some embodiments, a dog is determined to be at risk of GI disease if one or more values ​​in the fecal sample are different from the control values.

[0048] In some embodiments, a subject dog is determined to have no risk of GI disease if each of the fecal sample values ​​is similar to or identical to the control value, or to have a risk of GI disease if each of the fecal sample values ​​is different from the control value.

[0049] In some embodiments, a dog is determined to be at no risk of GI disease if its fecal sample has, compared to a control value, (i) a total short-chain fatty acid content at approximately the same or higher level, (ii) a C2 SCFA at approximately the same or higher level, (iii) a C3 SCFA at approximately the same or higher level, (iv) a C4 SCFA at approximately the same or higher level, (v) a pH at approximately the same or lower level, (vi) a fecal score at approximately the same or higher level, and (vii) an approximately the same or higher abundance of Bacteroidetes, Prevotellaceae, Ruminococcusceae, Bacteroidaceae, and Prevotella bacteria, and an approximately the same or lower abundance of Enterobacteriaceae. In some embodiments, a dog is determined to be at risk of GI disease if its fecal sample has, compared to a control value, (i) a lower total content of short-chain fatty acids, (ii) a lower level of C2 SCFA, (iii) a lower level of C3 SCFA, (iv) a lower level of C4 SCFA, (v) a higher pH, (vi) a lower fecal score, (vii) a lower abundance of Bacteroidetes, Prevotellaceae, Ruminococcusceae, Bacteroidaceae, and Prevotella bacteria, and a higher abundance of Enterobacteriaceae bacteria.

[0050] In some embodiments, a dog is determined to be at no risk of GI disease if, compared to a control value, its fecal sample has: (i) a total short-chain fatty acid content at approximately the same or higher level; (ii) a C2 SCFA at approximately the same or higher level; (iii) a C3 SCFA at approximately the same or higher level; (iv) a C4 SCFA at approximately the same or higher level; (v) a pH at approximately the same or lower level; (vi) a moisture content at approximately the same or lower level; (vii) a fecal score at approximately the same or higher level; and (viii) an abundance of one or more bacteria from Prevotellaceae, Ruminococcaceae, and Bacteroidaceae, and an abundance of at least one or both of Enterobacteriaceae and Bacteroidetes other than Prevotellaceae and Bacteroidaceae at approximately the same or lower level. In some embodiments, a dog is determined to be at risk of GI disease if its fecal sample, compared to a control value, has at least one of the following: (i) a lower total content of short-chain fatty acids, (ii) a lower level of C2 SCFA, (iii) a lower level of C3 SCFA, (iv) a lower level of C4 SCFA, (v) a higher pH, (vi) more moisture, (vii) a lower fecal score, and (viii) one or more of the following: a lower abundance of one or more bacteria from Prevotellaceae, Ruminococcaceae, and Bacteroidaceae, and a higher abundance of one or both of Enterobacteriaceae and Bacteroidetes other than Prevotellaceae and Bacteroidaceae.

[0051] In some embodiments, the step of identifying a subject dog as being at risk or not at risk of gastrointestinal (GI) disease includes (a) analyzing a fecal sample from the subject dog to determine the values ​​of the fecal sample for one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (vii) bacterial abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes.

[0052] In some embodiments, a dog is determined to be free from the risk of GI disease if the values ​​of the fecal sample include (i) the total content of short-chain fatty acids (SCFA) found in dogs without risk in the examples herein, (ii) the level of C2 SCFA found in dogs without risk in the examples herein, (iii) the level of C3 SCFA found in dogs without risk in the examples herein, (iv) the level of C4 SCFA found in dogs without risk in the examples herein, (v) the pH found in dogs without risk in the examples herein, (vi) the moisture content found in dogs without risk in the examples herein, (vii) the fecal score found in dogs without risk in the examples herein, and (vii) the abundance of one or more bacteria from among Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes found in dogs without risk in the examples herein. In some embodiments, the values ​​of the fecal sample include each of (i) to (vii). In some embodiments, the values ​​of the fecal sample include the abundance of each of (i) to (vii), as well as the abundance of each of the bacteria Prevotellaceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes.

[0053] In some embodiments, a dog is determined to be at risk of GI disease if its fecal sample contains one or more of the following values: (i) a total short-chain fatty acid content (SCFA) lower than that observed in dogs without risk in the examples herein; (ii) a lower level of C2 SCFA than that observed in dogs without risk in the examples herein; (iii) a lower level of C3 SCFA than that observed in dogs without risk in the examples herein; and (iv) a lower level of C4 SCFA than that observed in dogs without risk in the examples herein. SCFA, (v) pH higher than that observed in dogs without risk in the examples herein, (vi) moisture higher than that observed in dogs without risk in the examples herein, (vii) fecal score lower than that observed in dogs without risk in the examples herein, and (viii) one or more of the following: a lower abundance of one or more bacteria from Prevotellaceae, Ruminococcaceae, and Bacteroidetes, and a higher abundance of one or both of Enterobacteriaceae and Bacteroidetes other than Prevotellaceae and Bacteroidetes, than that observed in dogs without risk in the examples herein. In some embodiments, the values ​​of the fecal sample include each of (i) to (vii). In some embodiments, the values ​​of the fecal sample include the abundance of each of (i) to (vii), as well as the abundance of each of the bacteria Prevotellaceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes.

[0054] In some embodiments, the GI disease relief diet comprises a dog food composition. In some embodiments, the dog food composition comprises one or more of quinoa, amaranth, eggshell membrane, and dietary fiber. In some embodiments, the composition comprises each of quinoa, amaranth, eggshell membrane, and dietary fiber. In some embodiments, the composition comprises about 1% to about 10% by weight of quinoa, about 1% to about 10% by weight of amaranth, about 0.5% to about 8% by weight of eggshell membrane, and about 2% to about 20% by weight of dietary fiber. In some embodiments, the dietary fiber comprises insoluble fiber and soluble fiber. In some embodiments, the dietary fiber comprises i) about 1% to about 18% by weight of insoluble fiber, and ii) about 0.5% to about 8% by weight of soluble fiber. The weight percentages are based on the total weight of the dog food composition on a dry material basis.

[0055] In some embodiments, the dog food composition further comprises about 0.1% to about 10% by weight of fatty acids. In some embodiments, the fatty acids comprise polyunsaturated fatty acids. In some embodiments, the polyunsaturated fatty acids comprise omega-3 fatty acids, omega-6 fatty acids, or a combination of two or more thereof. In some embodiments, the polyunsaturated fatty acids comprise omega-3 fatty acids selected from linolenic acid, stearidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid, or docosahexaenoic acid, or a combination of two or more thereof. In some embodiments, the omega-3 fatty acids comprise alpha-linolenic acid. In some embodiments, the omega-3 fatty acids comprise eicosapentaenoic acid or docosahexaenoic acid, or a combination thereof.

[0056] In some embodiments, the polyunsaturated fatty acids include omega-6 fatty acids selected from linolenic acid, calendic acid, eicosadienoic acid, arachidonic acid, docosadienoic acid, adrenaline, osbondic acid, tetracosatetraenoic acid, or tetracosapentaenoic acid, or two or more combinations thereof. In some embodiments, the polyunsaturated fatty acids include omega-6 fatty acids selected from linolenic acid, arachidonic acid, and two or more combinations thereof.

[0057] One embodiment includes an apparatus comprising an input, memory, a processor, and an output. In some embodiments, the apparatus is an electronic mobile device, a computer, or a mobile phone. The input is configured to receive the results of a method for identifying whether a subject dog is at risk or not of gastrointestinal (GI) disease. The memory includes processor-executable instructions for comparing the results with a control value and preparing a conclusion that there is or is not at risk based on the comparison, and for outputting the results. The processor is configured to execute the processor-executable instructions. The output is configured to display the conclusion. In some embodiments, the processor-executable instructions further include instructions that display a recommendation to administer a GI disease relief diet to the subject when it is concluded that the subject dog is at risk, the processor is further configured to execute the instructions and display the recommendation, and the output is further configured to display the recommendation. In some embodiments, the GI disease relief diet is a dog food composition described herein. In some embodiments, the dog food composition comprises one or more of quinoa, amaranth, eggshell membrane, and dietary fiber. In some embodiments, the composition comprises each of quinoa, amaranth, eggshell membrane, and dietary fiber. In some embodiments, the composition comprises about 1% to about 10% by weight of quinoa, about 1% to about 10% by weight of amaranth, about 0.5% to about 8% by weight of eggshell membrane, and about 2% to about 20% by weight of dietary fiber. In some embodiments, the dietary fiber comprises insoluble fiber and soluble fiber. In some embodiments, the dietary fiber comprises i) about 1% to about 18% by weight of insoluble fiber, and ii) about 0.5% to about 8% by weight of soluble fiber. The weight percentages are based on the total weight of the dog food composition on a dry matter basis. In some embodiments, any method for identifying dogs at risk or not at risk of digestive (GI) disease is any one of the methods described herein.

[0058] Certain aspects of further embodiments of this specification are as follows. These aspects are further embodiments of this specification. The above embodiments are not necessarily limited to one or more elements of any one of the following embodiments. However, further embodiments of this specification include providing any one or more elements of any one of the following embodiments in one of the above embodiments, or providing any one or more elements of any one of the following embodiments in place of one or more elements in one of the above embodiments.

[0059] According to the first aspect, a method for identifying whether a subject dog is at risk of or not having a gastrointestinal (GI) disease, (a) Analyze fecal samples from dogs for at least three of the following: pH, total short-chain fatty acid content (SCFA), and C2 SCFA, C3 SFFA, C4 SCFA, C5 SCFA, and / or C6 SCFA content, fecal score, moisture content, and the relative abundance of phylum bacteroidetes (excluding Prevotella seaae, F3:f_bacteroidaseae and F4:f_prevotella seaae), ruminococca seaae, Enterobacteria seaae, Clostridiales and / or Prevotella, and any other bacterial phylum, family, genus, or species. (b) This includes comparing the value obtained from (a) with a control value obtained from a single dog, or a pooled control value from multiple dogs that have been medically determined to be free from the risk of GI disease, The aforementioned fecal sample (i) pH approximately equal to or lower than the control value, and total SFCA at approximately equal to or higher than the control value, (ii) A method is provided for identifying a subject dog as being at no risk of GI disease if, when measured, it has a moisture content and / or relative abundance of Enterobacteria seaae at approximately the same or lower level as the control value, as well as C3 SCFA, C2 SFCA, C4 SCFA, fecal score at approximately the same or higher level as the control value, and relative abundance of Filam bacteroidetes, luminococca seaae, Clostridiales and / or Prevotella, excluding Prevotella seaae, F3:f_bacteroida seaae and F4:f_prevotella seaae.

[0060] The high diversity of the gut microbiome is also generally associated with dogs that are not at risk, which can determine the presence of other bacteria.

[0061] For the purposes of this embodiment, the term “three or more of” specifically includes four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, or fourteen or more of.

[0062] In some embodiments of this model, the "relative abundance" of bacteria can be determined as described in Examples 3 and 4.

[0063] For the purposes of this embodiment, the term “approximately” means having a variance of 10% or less, unless otherwise stated.

[0064] For the purposes of this embodiment, the term “control values ​​obtained from one dog, or pooled control values ​​from multiple dogs medically determined to be free from the risk of GI disease” may be from parallel experiments, or more likely from archived values ​​from prior experiments, including but not limited to the values ​​herein.

[0065] For the purposes of this embodiment, the term “risk-free” generally means as determined by a licensed veterinarian.

[0066] A kit is provided for identifying whether a subject dog is at risk of or not of GI disease, comprising: a reagent for determining the pH of a fecal sample; one or more reagents for measuring volatile carboxylic acids in the fecal sample; and one or more reagents for quantitatively identifying one or more of the following bacteria in the fecal sample: Prevotella seaae, F3:f_bacteroidetes seaae and F4:f_prevotella seaae, luminococcus seaae, Enterobacteria seaae, Clostridiales, Prevotella, and any other bacteria.

[0067] For the purposes of this embodiment, the term “reagents for determining pH” includes, but is not limited to, pH meters, pH test sticks, liquid pH indicators, metal electrode devices, glass electrode devices, semiconductor sensors, hydrogen electrode devices, quinhydrone electrode devices, ISFET (ion-sensitive field electrode transistor) electrodes, and antimony electrode devices.

[0068] For the purposes of this embodiment, the term “one or more reagents for measuring volatile carboxylic acids” includes, but is not limited to, those taught in Example 2 herein, bromocresol green reagent, gas chromatography / mass spectrometry, and high-performance liquid chromatography / mass spectrometry.

[0069] For the purposes of this embodiment, "one or more reagents for quantitatively identifying one or more of the following bacteria in a fecal sample, excluding Prevotella seaae, F3:f_bacteroidetes seaae and F4:f_prevotella seaae, luminococcus seaae, Enterobacteria seaae, Clostridiares, Prevotella, and any other bacteria" include, but are not limited to, 16S amplicon sequencing techniques, shotgun metagenomics techniques including any available sequencing platform (such as Illumina, 454 sequencing, nanopore, and PacBio), multiplex qPCR methods (or more) based on designing specific primers for individual components at the phylum, family, genus, and species levels, and commonly used polymerase chain reaction (PCR) reagents using aptamers.

[0070] Another aspect of the present invention provides a method for preventing or treating GI disease, comprising: identifying the GI status of a dog, i.e., identifying whether the dog is at risk of digestive (GI) disease; performing the method according to the first aspect of the present invention using a sample taken from the dog; and, if it is determined that the dog is at risk of GI disease, providing the dog with the GI disease relief diet described herein. Generally, dog food is a GI disease relief diet, tends to be high in total dietary fiber (at least 2% to about 20% by weight), has a useful soluble fiber to insoluble fiber ratio (e.g., about 1:2 to about 1:18), is low in fat (e.g., 10% or less of total calories, or 20% or less by weight), and contains polyunsaturated fats, omega-3 fatty acids, and omega-6 fatty acids.

[0071] In some embodiments, the GI disease relief diet is a dog food composition comprising a release-controlled pet food composition comprising a base material containing a fiber component including a highly soluble fiber source and a low soluble fiber source, as well as a polyphenol source, wherein the base material is configured to deliver the polyphenol source to the lower digestive (GI) tract of the mammal after it has been ingested by the mammal.

[0072] In some embodiments, the weight ratio of highly soluble fiber sources to low-soluble fiber sources in a GI disease relief diet is approximately 1:20 to 1:1. In some embodiments, the weight ratio of highly soluble fiber sources to low-soluble fiber sources is approximately 1:15 to 1:2. In other embodiments, the weight ratio of highly soluble fiber sources to low-soluble fiber sources is approximately 1:10 to 1:3. In yet another embodiment, the weight ratio of highly soluble fiber sources to low-soluble fiber sources is approximately 1:5 to 1:3. In yet another embodiment, the weight ratio of highly soluble fiber sources to low-soluble fiber sources is approximately 1:4.

[0073] In some embodiments, the highly soluble fiber source includes oat bran, buckwheat cracks, pea bran, barley, tomato meal, citrus pulp, beet pulp, and two or more combinations thereof. In some embodiments, the low soluble fiber source includes cellulosic materials, pecan fiber, or combinations thereof.

[0074] In some embodiments, the polyphenol source is of food or plant origin. In some embodiments, the food-derived polyphenol source includes polyphenols derived from fruits or vegetables.

[0075] In some embodiments, the polyphenol source comprises flavonoids or phenolic acids. In certain embodiments, the polyphenol source provides polyphenols selected from dehydrooxyrosmarinic acid, coumaroyl nepitrin, eupaphorin, carnosol, scutellarin, kaempferol, rosmarinic acid, rosmanol, silcimaritin, luteolin, 6-methoxy-luteolin, 7-epirothmannol, quercetin, catechin, hesperidin, cyanidin, and two or more combinations thereof.

[0076] In some embodiments, the GI disease relief diet further comprises a source of hydrolyzed animal or plant protein containing an amino acid profile. In some embodiments, the source of hydrolyzed animal or plant protein includes chicken liver. In other embodiments, the source of hydrolyzed animal or plant protein is present in an active content of about 25 to about 45% by weight.

[0077] In some embodiments, the GI disease relief diet further comprises high-docosahexaenoic acid fish oil. In other embodiments, the active content of high-docosahexaenoic acid fish oil is approximately 0.5 to approximately 2.5% by weight.

[0078] In some embodiments, the fibrous component comprises pecan husks, flaxseed, citrus pulp, beet pulp, cranberry pulp, or a combination of two or more thereof. In some embodiments, the GI disease relief diet comprises about 1% to about 10% by weight of pecan husks, about 1% to about 5% by weight of flaxseed, about 1% to about 5% by weight of citrus pulp, about 1% to about 5% by weight of beet pulp, and about 0.1% to about 2% by weight of cranberry pulp. In other embodiments, the GI disease relief diet comprises about 2.5% to about 8% by weight of pecan husks, about 2% to about 4% by weight of flaxseed, about 2% to about 3% by weight of citrus pulp, about 2% to about 3% by weight of beet pulp, and about 0.5% to about 1.5% by weight of cranberry pulp. In certain embodiments, the GI disease relief diet comprises approximately 7% by weight of pecan shells, approximately 3% by weight of flaxseed, approximately 2.5% by weight of citrus pulp, approximately 2.5% by weight of beet pulp, and approximately 1% by weight of cranberry pulp. Further embodiments provide a GI disease relief diet comprising approximately 3.5% by weight of pecan shells, approximately 1.5% by weight of flaxseed, approximately 1.25% by weight of citrus pulp, approximately 1.25% by weight of beet pulp, and approximately 0.5% by weight of cranberry pulp.

[0079] In another embodiment, the pet food composition further comprises about 0.1 to about 10% by weight of fatty acids.

[0080] According to the third embodiment, the fatty acids of the dog food composition of the first embodiment include polyunsaturated fatty acids.

[0081] According to the fourth aspect, the polyunsaturated fatty acids of the dog food composition of the third aspect include omega-3 fatty acids, omega-6 fatty acids, or a combination of two or more thereof.

[0082] According to the fifth embodiment, the polyunsaturated fatty acids of the dog food composition of the fourth embodiment include omega-3 fatty acids selected from linolenic acid, stearidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, and two or more combinations thereof.

[0083] According to the sixth aspect, the omega-3 fatty acid of the dog food composition of the fourth or fifth aspect includes alpha-linolenic acid.

[0084] According to the seventh embodiment, the omega-3 fatty acid of one of the dog food compositions from the fourth to sixth embodiments includes eicosapentaenoic acid, docosahexaenoic acid, or a combination thereof.

[0085] According to the eighth aspect, the polyunsaturated fatty acids of one of the dog food compositions from the fourth to seventh aspects include linolenic acid, calendic acid, eicosadienoic acid, arachidonic acid, docosadienoic acid, adrenaline, osbondic acid, tetracosatetraenoic acid, tetracosapentaenoic acid, and omega-6 fatty acids selected from two or more combinations thereof.

[0086] According to the ninth aspect, the polyunsaturated fatty acids of the dog food composition according to the fourth or eighth aspect include omega-6 fatty acids selected from linolenic acid, arachidonic acid, and two or more combinations thereof.

[0087] According to the tenth aspect, the dog food composition according to the eighth or ninth aspect has a weight ratio of linolenic acid to arachidonic acid of about 12:1 to about 50:1, preferably about 15:1 to about 40:1, preferably about 18:1 to about 35:1, preferably about 20:1 to about 30:1, preferably about 22:1 to about 28:1, or preferably about 26:1 to about 28:1.

[0088] According to the eleventh aspect, one of the dog food compositions from the fourth to the tenth aspects has a weight ratio of omega-3 fatty acids to omega-6 fatty acids of about 0.5:1 to about 7:1, preferably about 0.5:1 to about 3:1, preferably about 0.5:1 to about 2.5:1, preferably about 0.5:1 to about 2:1, preferably about 0.5:1 to about 1.5:1, preferably about 0.5:1 to about 1:1, about 0.6:1 to about 7:1, preferably about 0.6:1 to about 3:1, preferably about 0.6:1 to about 2.5:1, preferably about 0.6:1 to about 2:1, preferably about 0.6:1 to about 1.5:1, or preferably about 0.6:1 to about 1:1.

[0089] According to the twelfth aspect, a dog food composition according to any of the above-described aspects is 10% by weight of fat or less Approximately 10% to 40% by weight of protein, and It contains approximately 2% to 20% by weight of dietary fiber, and the dietary fiber is i) Approximately 1% to 18% by weight of insoluble fibers, and ii) Contains approximately 0.5% to 8% by weight of soluble fibers.

[0090] According to the thirteenth aspect, the dog food composition according to the twelfth aspect has a weight ratio of soluble fiber to insoluble fiber of about 1:20 to about 10:1, or about 1:18 to about 5:1, or about 1:18 to about 2:1, or about 1:18 to about 1:1, or about 1:18 to about 1:2.

[0091] According to the fourteenth aspect, the dog food composition of any of the aforementioned aspects comprises a core and a covering disposed on the core, wherein the core comprises lysine, quinoa, and amaranth, and the covering comprises eggshell membrane.

[0092] According to the fifteenth aspect, Alpha-amino acids, One or more ancient grains, A dog food composition containing eggshell membrane, The dog food composition is Approximately 1% to 8% by weight of omega-3 fatty acids, It contains approximately 1% to 15% by weight of omega-6 fatty acids. A dog food composition is provided, wherein the food composition has a weight ratio of omega-3 fatty acids to omega-6 fatty acids of approximately 0.5:1 to approximately 7:1, and all weight percentages are based on the total weight of the dog food composition on a dry matter basis.

[0093] According to the sixteenth aspect, the alpha-amino acid of the dog food composition according to the fifteenth aspect contains an alpha-carboxylic acid group.

[0094] According to the seventeenth aspect, the alpha-amino acid of the dog food composition according to the fifteenth or sixteenth aspect includes lysine.

[0095] According to the eighteenth aspect, one of the dog food compositions from the fifteenth to seventeenth aspects is one or more ancient grains selected from spelt, khorasan wheat, einkorn, emmer, millet, barley, teff, sorghum, quinoa, amaranth, buckwheat, chia, and two or more combinations thereof.

[0096] According to the nineteenth aspect, the dog food composition according to the eighteenth aspect comprises one or more ancient grains including amaranth.

[0097] According to the twentieth aspect, one of the dog food compositions from the fifteenth to nineteenth aspects comprises at least two ancient grains.

[0098] According to the 21st aspect, one of the dog food compositions from the 15th to the 20th aspects has a weight ratio of omega-3 fatty acids to omega-6 fatty acids of about 0.5:1 to about 3:1, preferably about 0.5:1 to about 2.5:1, preferably about 0.5:1 to about 2:1, preferably about 0.5:1 to about 1.5:1, or preferably about 0.5:1 to about 1:1.

[0099] According to the 22nd aspect, one of the dog food compositions from the 15th to 21st aspects is a dog food composition in which the omega-3 fatty acid is selected from linolenic acid, stearidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, or a combination of two or more thereof.

[0100] According to the 23rd aspect, one of the dog food compositions from the 15th to 22nd aspects is one in which the omega-6 fatty acid is selected from linolenic acid, calendic acid, eicosadienoic acid, arachidonic acid, docosadienoic acid, adrenaline, osbondic acid, tetracosatetraenoic acid, tetracosapentaenoic acid, and two or more combinations thereof.

[0101] According to the twenty-fourth aspect, (a) core, Quinoa, The core, including amaranth, (b) A covering placed on the core, A dog food composition comprising a coating containing eggshell membrane, The dog food composition has a value of major nutrients containing approximately 2% to 10% by weight of dietary fiber, and the dietary fiber is Approximately 1% to 9% by weight of insoluble fibers, and It contains approximately 0.2% to 4% by weight of soluble fibers. A dog food composition is provided, wherein the food composition has a weight ratio of soluble fiber to insoluble fiber of approximately 1:20 to approximately 2:1, and all weight percentages are based on the total weight of the dog food composition on a dry matter basis.

[0102] According to the 26th aspect, the value of major nutrients in the dog food composition according to either the 24th or 25th aspect further includes fatty acids in an amount of about 0.1% to about 10% by weight.

[0103] According to the 27th aspect, the fatty acids in the dog food composition according to the 26th aspect include polyunsaturated fatty acids.

[0104] According to the 28th aspect, the polyunsaturated fatty acids of the dog food composition according to the 27th aspect include omega-3 fatty acids, omega-6 fatty acids, or a combination of two or more thereof.

[0105] According to the 29th aspect, the polyunsaturated fatty acids of the dog food composition according to the 28th aspect include omega-3 fatty acids selected from linolenic acid, stearidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, and two or more combinations thereof.

[0106] According to the 30th aspect, the omega-3 fatty acid of the dog food composition according to the 28th or 29th aspect includes alpha-linolenic acid.

[0107] According to the 31st aspect, the omega-3 fatty acid of one of the dog food compositions from the 28th to the 30th aspects includes eicosapentaenoic acid, docosahexaenoic acid, or a combination thereof.

[0108] According to the 32nd aspect, a dog food composition according to one of the 28th to 30th aspects is a dog food composition according to one of the 24th to 26th aspects, wherein the polyunsaturated fatty acids include omega-6 fatty acids selected from linolenic acid, calendic acid, eicosadienoic acid, arachidonic acid, docosadienoic acid, adrenaline, osbondic acid, tetracosatetraenoic acid, tetracosapentaenoic acid, and two or more combinations thereof.

[0109] According to the 33rd aspect, the dog food composition according to the 28th or 32nd aspect comprises omega-6 fatty acids selected from linolenic acid, arachidonic acid, and two or more combinations thereof, as polyunsaturated fatty acids.

[0110] According to the 34th aspect, the dog food composition according to the 32nd or 33rd aspect has a weight ratio of linolenic acid to arachidonic acid of about 12:1 to about 50:1, preferably about 15:1 to about 40:1, preferably about 18:1 to about 35:1, preferably about 20:1 to about 30:1, or preferably about 22:1 to about 28:1.

[0111] According to the 35th aspect, one of the dog food compositions from the 28th to 34th aspects has a weight ratio of omega-3 fatty acids to omega-6 fatty acids of about 0.5:1 to about 7:1, preferably about 0.5:1 to about 3:1, preferably about 0.5:1 to about 2.5:1, preferably about 0.5:1 to about 2:1, preferably about 0.5:1 to about 1.5:1, preferably about 0.5:1 to about 1:1, preferably about 0.6:1 to about 7:1, preferably about 0.6:1 to about 3:1, preferably about 0.6:1 to about 2.5:1, preferably about 0.6:1 to about 2:1, preferably about 0.6:1 to about 1.5:1, or preferably about 0.6:1 to about 1:1.

[0112] According to the 36th aspect, one of the dog food compositions from the 28th to 35th aspects further comprises alpha-amino acids.

[0113] According to the 37th aspect, the dog food composition according to the 36th aspect contains an alpha-amino acid with an alpha-carboxylic acid group.

[0114] According to the 38th aspect, the alpha-amino acid of the dog food composition according to the 37th aspect includes lysine.

[0115] According to the 39th aspect, the dog food composition of any of the aforementioned aspects is in the form of a dog treat and / or supplement adapted for combination with dog food.

[0116] According to the fortieth aspect, A certain amount of quinoa, A certain amount of amaranth, and A certain amount of eggshell membrane, A dog food composition containing approximately 2% to approximately 10% by weight of dietary fiber, wherein the dietary fiber is Approximately 1% to 9% by weight of insoluble fibers, and It contains approximately 0.2% to 4% by weight of soluble fibers. The dog food composition has a weight ratio of quinoa to amaranth of approximately 1:12 to approximately 12:1, optionally approximately 1:10 to approximately 10:1, optionally approximately 1:8 to approximately 8:1, optionally approximately 1:7 to approximately 7:1, optionally approximately 1:6 to approximately 6:1, optionally approximately 1:5 to approximately 5:1, optionally approximately 1:4 to approximately 4:1, optionally approximately 1:3 to approximately 3:1, or optionally approximately 1:2 to approximately 2:1. The dog food composition has a weight ratio of quinoa to eggshell membrane of approximately 1:10 to approximately 12:1, optionally approximately 1:8 to approximately 10:1, optionally approximately 1:6 to approximately 8:1, optionally approximately 1:5 to approximately 7:1, optionally approximately 1:4 to approximately 6:1, optionally approximately 1:3 to approximately 5:1, optionally approximately 1:2 to approximately 4:1, optionally approximately 1:1 to approximately 3:1, or optionally approximately 1:1 to approximately 2:1. The dog food composition is provided, having a weight ratio of amaranth to eggshell membrane of approximately 1:10 to approximately 12:1, optionally approximately 1:8 to approximately 10:1, optionally approximately 1:6 to approximately 8:1, optionally approximately 1:5 to approximately 7:1, optionally approximately 1:4 to approximately 6:1, optionally approximately 1:3 to approximately 5:1, optionally approximately 1:2 to approximately 4:1, optionally approximately 1:1 to approximately 3:1, or optionally approximately 1:1 to approximately 2:1.

[0117] According to the forty-first aspect, the dog food composition according to the fortyth aspect has a weight ratio of quinoa, amaranth, and eggshell membrane (quinoa:amaranth:eggshell membrane) of about 1:10:8 to about 10:10:1.

[0118] According to the forty-second aspect, the dog food composition according to the forty-fourth or forty-fifth aspect is: Approximately 1% to 10% by weight of quinoa; Approximately 1% to 10% by weight of amaranth, Approximately 0.5% to 8% by weight of eggshell membrane, It contains approximately 2% to 10% by weight of dietary fiber, and this dietary fiber is Approximately 1% to 9% by weight of insoluble fibers, and It contains approximately 0.2% to 4% by weight of soluble fibers. All weight percentages are based on the total weight of the dog food composition on a dry matter basis.

[0119] The following features may be applied to any or all aspects of this embodiment of the present invention.

[0120] To the extent that food and food components contain water / moisture, the dry substance represents everything in the sample other than water, such as proteins, fiber, fats, carbohydrates, and minerals. The weight of the dry substance is the total weight minus the weight of the water. Those skilled in the art will readily recognize and understand the amount of nutrients and proportions expressed as the quantity, weight, and percentage of the dry substance.

[0121] Daily dry matter intake is calculated as the total daily nutrient intake excluding all water. For example, the amount of a component corresponding to a specific percentage of daily nutrient intake represents the amount of that component in dry form (likewise excluding all water) relative to the total amount of dry matter consumed in a day (i.e., excluding all water).

[0122] "Daily nutrient intake" and "total daily nutrient intake" refer to the amount of dry matter consumed per day. In other words, the weight of water is not included in the calculation of daily nutrient consumption. To calculate the proportion of each component in the total daily intake based on dry matter, water is removed from the total intake to obtain the total daily dry matter intake, and the proportion of each component is calculated based on the amount of each component present as dry matter.

[0123] As used herein, “component” refers to any element of a dog food composition. The term “nutrient” refers to a substance that provides nutrition and therefore has nutritional value. In some cases, a component may contain two or more “nutrients,” for example, the composition may contain corn, which contains important nutrients including both protein and carbohydrates.

[0124] A dog food composition according to a particular aspect of the present invention comprises about 0.1% to about 5% by weight of alpha-amino acids, about 1% to about 15% by weight of one or more ancient grains (e.g., about 1% to about 10% by weight of quinoa and about 1% to about 10% by weight of amaranth), and about 0.5% to about 8% by weight of eggshell membrane, all weight percentages being based on the total weight of the dog food composition on a dry weight basis. In some aspects, the dog food composition may contain one or more fatty acids, such as polyunsaturated fatty acids. For example, the dog food composition may contain polyunsaturated fatty acids selected from omega-3 fatty acids, omega-6 fatty acids, or combinations of two or more thereof. The dog food composition may be formulated to have omega-3 fatty acids and omega-6 fatty acids in a weight ratio of preferably about 0.5:1 to about 7:1. In some embodiments, the dog food composition may be formulated to control the weight ratio of linolenic acid to arachidonic acid to about 12:1 to about 50:1.

[0125] The dog food composition may also have macronutrient values ​​including approximately 2% to 20% by weight of dietary fiber, which consists of approximately 1% to 18% by weight of insoluble fiber and approximately 0.5% to 4% by weight of soluble fiber. Furthermore, the dog food composition may have a weight ratio of soluble fiber to insoluble fiber of approximately 1:20 to 5:1, and all weight percentages are based on the total weight of the dog food composition on a dry matter basis.

[0126] In a further embodiment, the dog food composition is in the form of having a core and a coating placed on the core. The core typically contains lysine and amaranth, while the coating contains eggshell membrane. Although not limited to any particular theory, it is thought that including eggshell membrane in the coating may allow for the maintenance of higher concentrations of bioactive peptides in the eggshell membrane.

[0127] Appropriate ingredients, such as those listed below, may be included in or excluded from the formulation of the dog food composition, depending on the specific combination of other ingredients and the form of the dog food composition. In some embodiments, the dog food compositions disclosed herein may be in the form of standalone dog food, dog food supplements, dog treats, or similar.

[0128] Dog food compositions typically contain one or more ancient grains. As used herein, ancient grains refer to grains such as wheat, millet, barley, teff, oats, and sorghum, as well as pseudo-grains such as amaranth, quinoa, buckwheat, and chia. In some embodiments, dog food compositions contain two or more ancient grains, three or more ancient grains, four or more ancient grains, or five or more ancient grains. The ancient grains may optionally be selected from millet, barley, teff, oats, sorghum, amaranth, buckwheat, chia, and two or more combinations thereof. Preferably, the ancient grains are selected from quinoa, amaranth, buckwheat, bulgur, sorghum, and two or more combinations thereof. In some cases, dog food compositions may contain at least two ancient grains selected from quinoa, amaranth, buckwheat, bulgur, sorghum, and two or more combinations thereof. For example, the dog food composition may include quinoa and amaranth.

[0129] Additionally, or alternatively, the dog food composition may contain a first ancient grain (e.g., quinoa) and a second ancient grain (e.g., amaranth) in a weight ratio of about 1:12 to about 12:1. In some cases, the dog food composition has a weight ratio of the first ancient grain to the second ancient grain of about 1:10 to about 10:1, about 1:8 to about 8:1, about 1:7 to about 7:1, about 1:6 to about 6:1, about 1:5 to about 5:1, about 1:4 to about 4:1, about 1:3 to about 3:1, or about 1:2 to about 2:1, or any range or partial range thereof. In at least one embodiment, the weight ratio of the first ancient grain (e.g., quinoa) to the second ancient grain (e.g., amaranth) is about 1:1 or 1:1.

[0130] One or more ancient grains may be included in the dog food composition in an amount ranging from approximately 1% to approximately 15% by weight, based on the total weight of the dog food composition on a dry matter basis. For example, one or more ancient grains may be present in amounts of approximately 1–15% by weight, approximately 1–12% by weight, approximately 1–10% by weight, approximately 1–8% by weight, approximately 1–6% by weight, approximately 1–4% by weight, approximately 1–3% by weight, approximately 2–15% by weight, approximately 2–12% by weight, approximately 2–10% by weight, approximately 2–8% by weight, approximately 2–6% by weight, approximately 2–4% by weight, approximately 2–3% by weight, approximately 3–15% by weight, approximately 3–12% by weight, approximately 3–10% by weight, approximately 3–8% by weight, approximately 3–6% by weight, approximately 3–4% by weight, approximately 4–15% by weight, approximately 4–12% by weight, approximately 4–10% by weight, approximately 4–8% by weight, or approximately 4–6% by weight (including these ranges and partial ranges). In at least one embodiment, the dog food composition contains about 2.5% by weight or 2.5% by weight of a first ancient grain such as quinoa, based on the total weight of the dog food composition on a dry matter basis. Additionally or alternatively, the dog food composition may contain about 2.5% by weight or 2.5% by weight of a second ancient grain such as amaranth, based on the total weight of the dog food composition on a dry matter basis.

[0131] Dog food compositions typically contain eggshell membrane. Eggshell membrane typically contains protein components, hyaluronic acid, and certain bioactive peptides that may be beneficial in the dog food compositions disclosed herein. Without limiting to any particular theory, dog food compositions containing eggshell membrane instead of eggs or dried eggs are thought to significantly reduce allergic reactions in dogs suffering from egg allergies. Eggshell membrane may be present in the dog food composition in an amount ranging from about 0.5% to about 8% by weight, based on the total weight of the dog food composition on a dry matter basis. For example, a dog food composition may contain eggshell membrane in amounts of approximately 0.5 to approximately 8% by weight, approximately 0.5 to approximately 7% by weight, approximately 0.5 to approximately 6% by weight, approximately 0.5 to approximately 5% by weight, approximately 0.5 to approximately 4% by weight, approximately 0.5 to approximately 3% by weight, approximately 0.5 to approximately 2% by weight, approximately 1 to approximately 8% by weight, approximately 1 to approximately 7% by weight, approximately 1 to approximately 6% by weight, approximately 1 to approximately 5% by weight, approximately 1 to approximately 4% by weight, approximately 1 to approximately 3% by weight, approximately 1 to approximately 2% by weight, approximately 2 to approximately 8% by weight, approximately 2 to approximately 7% by weight, approximately 2 to approximately 6% by weight, approximately 2 to approximately 5% by weight, approximately 2 to approximately 4% by weight, approximately 2 to approximately 3% by weight, approximately 3 to approximately 8% by weight, approximately 3 to approximately 7% by weight, approximately 3 to approximately 6% by weight, approximately 3 to approximately 5% by weight, or approximately 3 to approximately 4% by weight (including these ranges and partial ranges). In at least one embodiment, the dog food composition contains about 2% by weight or 2% by weight of eggshell membrane, based on the total weight of the dog food composition on a dry matter basis.

[0132] In some embodiments, the dog food composition is formulated to have a specific weight ratio of ancient grains to eggshell membrane. For example, the dog food composition may have a weight ratio of ancient grains to eggshell membrane of approximately 1:10 to approximately 24:1, approximately 1:8 to approximately 20:1, approximately 1:6 to approximately 16:1, approximately 1:5 to approximately 14:1, approximately 1:4 to approximately 12:1, approximately 1:3 to approximately 10:1, approximately 1:2 to approximately 8:1, approximately 1:1 to approximately 6:1, approximately 1:1 to approximately 4:1, or any range or partial range thereof. In at least one embodiment, the dog food composition comprises at least two ancient grains, the weight ratio of a first ancient grain (e.g., quinoa) to eggshell membrane being approximately 1:10 to 12:1, 1:8 to 10:1, 1:6 to 8:1, 1:5 to 7:1, 1:4 to 6:1, 1:3 to 5:1, 1:2 to 4:1, 1:1 to 6:1, 1:1 to 5:1, 1:1 to 4:1, 1:1 to 3:1, 1:1 to 2:1, or any range or partial range thereof. Additionally or alternatively, the dog food composition may have a weight ratio of a second ancient grain (e.g., amaranth) to eggshell membrane of approximately 1:10 to 12:1, 1:8 to 10:1, 1:6 to 8:1, 1:5 to 7:1, 1:4 to 6:1, 1:3 to 5:1, 1:2 to 4:1, 1:1 to 6:1, 1:1 to 5:1, 1:1 to 4:1, 1:1 to 3:1, 1:1 to 2:1, or any range or partial range thereof.

[0133] In some cases, the dog food composition is formulated to preferably have a specific weight ratio of quinoa, amaranth, and eggshell membrane (quinoa:amaranth:eggshell membrane) of about 1:10:8 to about 10:10:1. For example, the weight ratio of quinoa, amaranth, and eggshell membrane may be about 1:10:8 to about 10:10:1, about 1:5:4 to about 5:5:1, about 1:8:4 to about 10:2:4, about 1:4:2 to about 5:1:2, or any range or partial range thereof.

[0134] Optionally, the dog food composition may contain one or more alpha-amino acids. The alpha-amino acids may contain an alpha-carboxylic acid group. In some embodiments, one or more alpha-amino acids include lysine. The alpha-amino acids may be present in the food composition in an amount ranging from about 0.1% to about 5% by weight, based on the total weight of the dog food composition on a dry matter basis. For example, dog food composition is based on the total weight of the dog food composition on a dry matter basis, and is approximately 0.1% to 5% by weight, approximately 0.1% to 4% by weight, approximately 0.1% to 3% by weight, approximately 0.1% to 2% by weight, approximately 0.1% to 1% by weight, approximately 0.2% to 5% by weight, approximately 0.2% to 4% by weight, approximately 0.2% to 3% by weight, approximately 0.2% to 2% by weight, approximately 0.2% to 1% by weight, approximately 0.5% to 5% by weight, approximately 0.5% to 4% by weight, approximately 0.5% to 3% by weight, approximately 0.5% to 2% by weight, approximately The dog food composition may contain one or more alpha-amino acids in amounts of 0.5% to about 1% by weight, about 0.75% to about 5% by weight, about 0.75% to about 4% by weight, about 0.75% to about 3% by weight, about 0.75% to about 2% by weight, about 0.75% to about 1% by weight, about 1% to about 5% by weight, about 1% to about 4% by weight, about 1% to about 3% by weight, about 1% to about 2% by weight, about 2% to about 5% by weight, about 2% to about 4% by weight, about 2% to about 3% by weight, about 3% to about 5% by weight, or any range or partial range thereof. In at least one embodiment, the dog food composition contains alpha-amino acids, preferably lysine, in amounts of about 2.5% by weight or 2.5% by weight, based on the total weight of the dog food composition on a dry matter basis.

[0135] Additional ingredients may be included, or excluded in some cases, including beef broth, dried brewer's yeast, eggs, egg products, split flax, 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 mussel, blue-lipped mussel. Mussel, methylsulfonylmethane (MSM), boron, boric acid, phytoestrogens, phytoandrogens, genistein, daidzein, L-carnitine, chromium picolinate, chromium tripicolinate, 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, and chlorine. Examples of such agents include denaturants, pharmacoagulants, astringents, emulsifiers, topical analgesics, aromatic compounds, humectants, opacifying agents (such as zinc oxide and titanium dioxide), defoaming agents (such as silicones), preservatives (butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), propyl gallate, benzalkonium chloride, EDTA, benzyl alcohol, potassium sorbate, parabens, and mixtures thereof), reducing agents, solvents, hydrotropes, solubilizers, suspending agents (non-surfactants), solvents, viscosity enhancers (aqueous and non-aqueous), scavengers, and / or keratolytic agents.

[0136] Dog food compositions are typically formulated to contain one or more fatty acids. Non-limiting examples of fatty acids include, but are not limited to, omega-3 fatty acids, omega-6 fatty acids, lauric acid, myristic acid, palmitic acid, palmitoleic acid, margaric acid, margaroleic acid, stearic acid, oleic acid, stearidonic acid, gadoleic acid, behenic acid, erucic acid, docosatetraic acid, and two or more combinations thereof. The fatty acids may be polyunsaturated fatty acids, such as omega-3 fatty acids, omega-6 fatty acids, or two or more combinations thereof. Non-limiting examples of omega-3 fatty acids include linolenic acid, stearidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, and combinations of two or more thereof. Dog food compositions may contain linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, or two or more combinations thereof. In at least one embodiment, the dog food composition comprises alpha-linolenic acid and / or gamma-linolenic acid. In a further embodiment, the dog food composition comprises one or more omega-3 fatty acids, including eicosapentaenoic acid, docosahexaenoic acid, or a combination thereof.

[0137] Additionally, or alternatively, the polyunsaturated fatty acids may include omega-6 fatty acids. Examples of omega-6 fatty acids include linolenic acid, calendic acid, eicosadienoic acid, arachidonic acid, docosadienoic acid, adrenaline, osbondic acid, tetracosatetraenoic acid, tetracosapentaenoic acid, or two or more combinations thereof. In some embodiments, the polyunsaturated fatty acids include omega-6 fatty acids selected from linolenic acid, arachidonic acid, and two or more combinations thereof. In at least one preferred embodiment, the dog food composition is formulated to have a weight ratio of linolenic acid to arachidonic acid of about 12:1 to about 50:1. In some cases, the weight ratio of linolenic acid to arachidonic acid is approximately 15:1 to 40:1, 15:1 to 35:1, 15:1 to 30:1, 15:1 to 28:1, 18:1 to 40:1, 18:1 to 35:1, 18:1 to 30:1, 18:1 to 28:1, 20:1 to 40:1, and 20 :1 to approximately 35:1, approximately 20:1 to approximately 30:1, approximately 20:1 to approximately 28:1, approximately 22:1 to approximately 40:1, approximately 22:1 to approximately 35:1, approximately 22:1 to approximately 30:1, approximately 2:1 to approximately 28:1, approximately 18:1 to approximately 35:1, approximately 20:1 to approximately 30:1, approximately 22:1 to approximately 28:1, or any range or subrange thereof.

[0138] Additionally, or alternatively, the dog food composition may be formulated to have a weight ratio of omega-3 fatty acids to omega-6 fatty acids of approximately 0.5:1 to approximately 7:1. In some embodiments, the dog food composition has a weight ratio of omega-3 fatty acids to omega-6 fatty acids of approximately 0.5:1 to approximately 6:1, approximately 0.5:1 to approximately 5:1, approximately 0.5:1 to approximately 4:1, approximately 0.5:1 to approximately 3:1, approximately 0.5:1 to approximately 2.5:1, approximately 0.5:1 to approximately 2:1, approximately 0.5:1 to approximately 1.5:1, or approximately 0.5:1 to approximately 1:1, approximately 1:1 to approximately 6:1, approximately 1:1 to approximately 5:1, approximately 1:1 to approximately 4:1, approximately 1:1 to approximately 3:1, approximately 1:1 to approximately 2.5:1, approximately 1:1 to approximately 2:1, approximately 1:1 to approximately 1.5:1, or approximately 1:1 to approximately 1:1 (including those ranges or partial ranges).

[0139] In some embodiments, the food composition further comprises one or more amino acids. One or more amino acids may be included in the dog food composition as free amino acids, or they may be supplied to the composition of the Disclosure by any number of component sources (e.g., crude protein). Examples of amino acids, but not limited to, include tryptophan, taurine, histidine, carnitine, carnosine, alanine, cysteine, arginine, methionine (including DL-methionine and L-methionine), tryptophan, lysine, asparagine, aspartate (aspartic acid), phenylalanine, valine, threonine, isoleucine, histidine, leucine, glycine, glutamine, taurine, tyrosine, homocysteine, ornithine, citrulline, glutamate (glutamic acid), proline, and / or serine.

[0140] One or more amino acids may include essential amino acids. Essential amino acids are those that cannot be newly synthesized or that organisms cannot synthesize in sufficient quantities, and therefore must be supplied through diet. Essential amino acids vary from species to species, depending on the organism's metabolism. For example, it is generally understood that the essential amino acids for dogs are phenylalanine, leucine, methionine, lysine, isoleucine, valine, threonine, tryptophan, histidine, and arginine.

[0141] Dog food compositions typically contain total dietary fiber in amounts ranging from approximately 2% to approximately 20% by weight, based on the total weight of the dog food composition on a dry matter basis. For example, total dietary fiber may be present in amounts of approximately 2–20% by weight, approximately 2–16% by weight, approximately 2–12% by weight, approximately 2–10% by weight, approximately 2–8% by weight, approximately 2–6% by weight, approximately 6–20% by weight, approximately 6–16% by weight, approximately 6–12% by weight, approximately 6–10% by weight, approximately 8–20% by weight, approximately 8–16% by weight, approximately 8–12% by weight, approximately 8–10% by weight, approximately 10–20% by weight, approximately 10–16% by weight, approximately 12–20% by weight, approximately 12–16% by weight, approximately 14–20% by weight, or approximately 14–16% by weight (including those ranges and partial ranges).

[0142] Dietary fiber refers to plant components that are resistant to digestion by animal digestive enzymes. Dietary fiber includes soluble fiber and insoluble fiber. Soluble fiber is resistant to digestion and absorption in the small intestine and is completely or partially fermented in the large intestine, and includes, for example, beet pulp, guar gum, chicory root, psyllium, pectin, blueberries, cranberries, pumpkin, apples, oats, legumes, citrus fruits, barley, or peas. Insoluble fiber can be supplied from any variety of component sources, including, for example, pecan hulls, buckwheat hulls, cellulose, whole wheat products, wheat oat, corn bran, flaxseed, grapes, celery, green beans, cauliflower, potato peels, fruit peels, vegetable peels, peanut shells, and soy fiber. Crude fiber includes, for example, the hulls of grains such as rice, corn, and beans, and other indigestible components contained in the cell walls and cellular contents of plants such as grains. The typical amount of crude fiber in the composition of the Disclosure may be about 0 to about 20% by weight, about 1 to about 20% by weight, about 1 to about 15% by weight, about 1 to about 10% by weight, about 1 to about 7% by weight, about 1 to about 5% by weight, about 1 to about 3% by weight, about 3 to about 20% by weight, about 3 to about 15% by weight, about 3 to about 10% by weight, about 3 to about 7% by weight, about 3 to about 5% by weight, about 5 to about 20% by weight, about 5 to about 15% by weight, about 5 to about 10% by weight, about 5 to about 7% by weight, about 7 to about 20% by weight, about 7 to about 15% by weight, or about 7 to about 10% by weight (including any range or partial range thereof).

[0143] The dog food composition may contain about 1% to about 9% by weight of insoluble fiber, based on the total weight of the dog food composition on a dry matter basis. For example, the amount of insoluble fiber present in the dog food composition may be about 1 to about 9% by weight, about 1 to about 8% by weight, about 1 to about 7% by weight, about 1 to about 6% by weight, about 1 to about 5% by weight, about 1 to about 4% by weight, about 1 to about 3% by weight, about 2 to about 9% by weight, about 2 to about 8% by weight, about 2 to about 7% by weight, about 2 to about 6% by weight, about 2 to about 5% by weight, about 2 to about 4% by weight, about 3 to about 9% by weight, about 3 to about 8% by weight, about 3 to about 7% by weight, about 3 to about 6% by weight, about 3 to about 5% by weight, about 4 to about 9% by weight, about 4 to about 8% by weight, about 4 to about 7% by weight, or about 4 to about 6% by weight (including those ranges and partial ranges).

[0144] The dog food composition may contain about 0.2% to about 4% by weight of soluble fiber, based on the total weight of the dog food composition on a dry matter basis. For example, the amount of soluble fiber present in the dog food composition may be about 0.2 to about 4% by weight, about 0.2 to about 3% by weight, about 0.2 to about 2% by weight, about 0.2 to about 1% by weight, about 0.5 to about 4% by weight, about 0.5 to about 3% by weight, about 0.5 to about 2% by weight, about 0.5 to about 1% by weight, about 0.75 to about 4% by weight, about 0.75 to about 3% by weight, about 0.75 to about 2% by weight, about 1 to about 4% by weight, about 1 to about 3% by weight, about 1 to about 2% by weight, about 2 to about 4% by weight, or about 2 to about 3% by weight (including those ranges and partial ranges).

[0145] The dog food composition may be formulated to have a weight ratio of soluble fiber to insoluble fiber of approximately 1:20 to approximately 10:1. For example, the dog food composition may have a weight ratio of soluble fiber to insoluble fiber of approximately 1:18 to approximately 5:1, or approximately 1:18 to approximately 2:1, or approximately 1:18 to approximately 1:1, or approximately 1:18 to approximately 1:2.

[0146] Dog food compositions typically contain protein and / or digestible crude protein. The term “protein” means a polypeptide, peptide, or polymer of amino acids. This term encompasses naturally occurring and non-naturally occurring (synthetic) polymers, as well as polymers in which artificial chemical mimics are substituted for one or more amino acids. The term also encompasses fragments, variants, and homologs having the same or substantially the same properties and performing the same or substantially the same functions as the original sequence. This term encompasses polymers of any length, including polymers containing approximately 2–1000, 4–800, 6–600, and 8–400 amino acids. Proteins may also include synthesized, as well as amino acid polymers isolated and purified from natural sources. Under certain embodiments, the terms “polypeptide,” “peptide,” or “protein” are used interchangeably.

[0147] "Digestive crude protein" is the portion of a protein that, after being digested by stomach enzymes, can be used or converted into free nitrogen (amino acids). In vitro measurement of digestible crude protein may be achieved by using stomach enzymes such as pepsin, digesting the sample, and measuring the free amino acids after digestion. In vivo measurement of digestible crude protein may be achieved by measuring the protein level in a feed / food sample, feeding the sample to an animal, and measuring the amount of nitrogen collected in the animal's feces.

[0148] The protein and / or digestible crude protein in the composition may be present in varying amounts or concentrations. In one embodiment, the protein may be present in an amount of about 10% to about 40% by weight, based on the total weight of the dog food composition on a dry matter basis. For example, the protein may be present in an amount of about 10 to about 40% by weight, about 10 to about 36% by weight, about 10 to about 32% by weight, about 10 to about 28% by weight, about 10 to about 24% by weight, about 10 to about 20% by weight, about 10 to about 18% by weight, about 12 to about 40% by weight, about 12 to about 36% by weight, about 12 to about 32% by weight, about 12 to about 28% by weight, about 12 to about 24% by weight, about 12 to about 20% by weight, and about 12 to about 18% by weight, based on the total weight of the dog food composition on a dry matter basis. It may be present in amounts of approximately 10% by weight, approximately 14–40% by weight, approximately 14–36% by weight, approximately 14–32% by weight, approximately 14–28% by weight, approximately 14–24% by weight, approximately 14–20% by weight, approximately 14–18% by weight, approximately 16–40% by weight, approximately 16–36% by weight, approximately 16–32% by weight, approximately 16–28% by weight, approximately 16–24% by weight, approximately 16–20% by weight, or approximately 16–18% by weight (including those ranges or partial ranges). In some embodiments, the dog food composition contains protein in amounts of approximately 10% by weight, approximately 15% by weight, approximately 20% by weight, approximately 25% by weight, approximately 30% by weight, approximately 35% by weight, approximately 40% by weight, or any range formed from thereto, based on the total weight of the dog food composition on a dry matter basis. In another embodiment, protein may be present in an amount of about 10–25% by weight, about 15–25% by weight, or about 15–20% by weight, based on the total weight of the dog food composition on a dry basis. In a particular embodiment, protein may be present in an amount of about 12–35% by weight, about 13–25% by weight, or about 15–25% by weight, based on the total weight of the dog food composition on a dry basis.

[0149] A portion of the protein in the composition may be digestible protein. For example, the composition may contain an amount of protein in which about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, about 95% or more, about 98% or more, or about 99% or more of the total protein is digestible protein. In some embodiments, for example, when the desired composition promotes weight loss, the portion of the protein that is digestible protein is about 60% or less, about 50% or less, about 40% or less, about 30% or less, about 20% or less, or about 10% or less, based on the total amount of protein in the dog food composition on a dry weight basis. In a further embodiment, the amount of digestible protein is about 10 to about 99% by weight, about 10 to about 95% by weight, about 10 to about 90% by weight, about 10 to about 70% by weight, about 10 to about 50% by weight, about 10 to about 30% by weight, about 30 to about 99% by weight, about 30 to about 95% by weight, about 30 to about 90% by weight, about 30 to about 70% by weight, about 30 to about 50% by weight, about 50 to about 99% by weight, about 50 to about 95% by weight, about 50 to about 90% by weight, about 50 to about 70% by weight, or about 70 to about 99% by weight, about 70 to about 95% by weight, or about 70 to about 90% by weight, including ranges within and partial ranges thereof.

[0150] Proteins may be supplied from any variety of sources well known to those skilled in the art, including plant sources, animal sources, microbial sources, or combinations thereof. For example, animal sources may include meat, meat by-products, seafood, dairy products, and eggs. Meat may include, for example, chicken, fish, and animal meat (including cattle, pigs, sheep, goats, and similar animals). Meat by-products may include, for example, lungs, kidneys, brains, livers, stomachs, and intestines. Plant proteins may include, for example, soybeans, cottonseed, and peanuts. Microbial sources may be used to synthesize amino acids (e.g., lysine, threonine, tryptophan, methionine) or intact proteins (such as proteins from the sources listed below).

[0151] Proteins or protein components may include, for example, chicken meal, poultry, poultry by-product meal, lamb, lamb meal, turkey, turkey meal, beef, beef by-products, offal, fishmeal, intestines, kangaroo, white fish, venison, soybean meal, soybean protein isolate, soybean protein concentrate, corn gluten meal, corn protein concentrate, distillation-dried grains and / or distillation-dried grain solutions, single-cell proteins (e.g., yeast, algae, and / or bacterial cultures), etc.

[0152] Proteins may remain intact, be completely hydrolyzed, or be partially hydrolyzed. The protein content of food can be determined by several methods well known to those skilled in the art, such as Method 988.05 published by the Association of Official Analytical Chemists in the Official Methods of Analysis ("OMA"). The amount of protein in the compositions disclosed herein may also be determined based on the amount of nitrogen in the composition according to methods known to those skilled in the art.

[0153] The compositions of the present invention may optionally contain lipids. The term “fat” generally refers to lipids or mixtures of lipids that may be generally solid or liquid at normal room temperature (e.g., 25°C) and pressure (e.g., 1 atmosphere). In some cases, fats may be viscous liquids or amorphous solids at standard room temperature and standard pressure. Fats may be completely incorporated into the food composition, deposited on the outside of the dog food composition, or a mixture of the two. In some embodiments, the dog food composition further comprises an effective amount of one or more substances selected from the group consisting of glucosamine, chondroitin, chondroitin sulfate, methylsulfonylmethane ("MSM"), creatine, antioxidants, Perna canaliculata, and mixtures thereof.

[0154] Lipids can be supplied by any of the various sources known to those skilled in the art, including meat, meat by-products, canola oil, fish oil (such as anchovy oil and menhedene oil), and plants. Examples of meat fat sources include poultry fat, turkey fat, lard, animal fat, and beef fat. Examples of vegetable fat sources include wheat, flax, rye, barley, rice, sorghum, corn, oats, millet, wheat germ, corn germ, soybeans, peanuts, and cottonseed, as well as these and other vegetable fat sources, such as corn oil, soybean oil, cottonseed oil, palm oil, palm kernel oil, linseed oil, canola oil, rapeseed oil, and / or oils derived from orestra.

[0155] In some cases, the fat in the composition is crude fat. Crude fat may be included in the composition in the amounts disclosed above with respect to total fat, such as 10% by weight, based on the total weight of the dog food composition on a dry matter basis. Alternatively, about 50% or more by weight, about 60% or more by weight, about 70% or more by weight, about 80% or more by weight, or about 90% or more by weight of total fat may be obtained from plant sources.

[0156] 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 for energy when hydrolyzed. By appropriately balancing the carbohydrate sources, those skilled in the art can manipulate the texture of the final product. For example, short-chain polysaccharides tend to be more viscous and gooey, while long-chain polysaccharides are less viscous and gooey than short-chain polysaccharides, and the desired texture of this hybrid food is achieved by long-chain polysaccharides and modified starches (such as natural or modified starches, cellulose, and the like). Carbohydrate mixtures may additionally contain any optional components such as added salts, spices, seasonings, vitamins, minerals, flavorings, colorings, and the like. The amount of any optional component depends at least in part to the nutritional requirements of different life stages of animals.

[0157] Carbohydrates include, but are not limited to, oat fiber, cellulose, peanut husks, beet pulp, parboiled rice, corn starch, corn gluten meal, cereals, and sorghum, and can be supplied from any variety of ingredient sources known to those skilled in the art. Examples of cereals that supply carbohydrates include, but are not limited to, wheat, durum, semolina, corn, barley, and rice. In certain embodiments, the carbohydrate component may consist of a mixture of one or more carbohydrate sources. The carbohydrate content of a food can be determined by any number of methods known to those skilled in the art.

[0158] Generally, the proportion of carbohydrates can be calculated as soluble non-nitrogenous matter ("NFE"), which can be calculated as follows: NFE = 100% - (Moisture %) - (Protein %) - (Lipid %) - (Ash %) - (Crude Fiber %). The amount of carbohydrates present in the composition (for example, calculated as NFE) may be about 10-90% by weight, about 10-70% by weight, about 10-50% by weight, about 10-40% by weight, about 10-30% by weight, about 10-20% by weight, about 20-90% by weight, about 20-70% by weight, about 20-50% by weight, about 20-40% by weight, about 30-90% by weight, about 30-70% by weight, about 30-50% by weight, about 30-40% by weight, about 50-90% by weight, about 50-70% by weight, or about 70-90% by weight, based on the total weight of the dog food composition on a dry matter basis.

[0159] In certain embodiments, the dog food composition contains moisture. Moisture may be present in varying amounts or concentrations. In one embodiment, moisture may be present in amounts ranging from about 3% to about 20% based on the total weight of the dog food composition. For example, moisture may be present in amounts ranging from about 3% by weight, about 5% by weight, about 5.5% by weight, about 6% by weight, about 6.5% by weight, about 7% by weight, about 7.5% by weight, about 8% by weight, about 8.5% by weight, about 9% by weight, about 9.5% by weight, about 10% by weight, about 10.5% by weight, about 11% by weight, about 11.5% by weight, about 12% by weight, about 12.5% ​​by weight, about 13% by weight, about 13.5% by weight, about 14% by weight, about 14.5% by weight, or about 15% by weight based on the total weight of the dog food composition. In another embodiment, moisture may be present in amounts of about 6% to about 12%, about 9% to about 13%, about 9% to about 11%, or about 9% to about 13% based on the total weight of the dog food composition. In certain embodiments, moisture may be present in amounts of about 5% to about 12%, about 6% to about 11%, or about 7% to about 10.0%, based on the total weight of the dog food composition. In further embodiments, moisture may be present in amounts of about 65% to about 85%, about 60% to about 80%, or about 60% to about 75%, based on the total weight of the dog food composition.

[0160] The dog food composition may contain one or more components and / or sources of glucose mimetic, carotenoids, and / or arginine and their derivatives. Sources of glucose mimetic may include glucose antagonists (including 2-deoxy-D-glucose, 5-thio-D-glucose, and 3-O-methylglucose), anhydro sugars (including 1,5-anhydro-D-glucitol, 2,5-anhydro-D-glucitol, and 2,5-anhydro-D-mannitol), mannoheptulose, and / or avocado extract containing mannoheptulose. Sources of carotenoids may include lutein, astaxanthin, zeaxanthin, bixin, lycopene, and / or beta-carotene. Sources of antioxidant components may include tocopherol (vitamin E), vitamin C, vitamin A, plant-derived materials, carotenoids (as described above), selenium, and / or CoQ10 (coenzyme Q10). In a preferred embodiment, the dog food composition contains high levels of arginine and its derivatives. The amount of arginine present in the composition may be about 0.01 to about 10.0% by weight, about 0.01 to about 5.0% by weight, about 0.01 to about 2.0% by weight, about 0.1 to about 10.0% by weight, about 0.1 to about 5.0% by weight, about 0.1 to about 2.0% by weight, about 0.5 to about 5.0% by weight, about 0.5 to about 2.0% by weight, about 1.5 to about 5.0% by weight, about 1.5 to about 2.0% by weight, about 0.5 to about 2.0% by weight, about 1.5 to about 5.0% by weight, about 1.5 to about 2.0% by weight, about 0.5% by weight, about 1.0% by weight, about 1.4% by weight, about 1.44% by weight, about 1.8% by weight, or about 2.0% by weight, based on the total weight of the composition on a dry weight basis. The arginine present in the composition may be L-arginine, D-arginine, or a mixture thereof.

[0161] The dog food compositions disclosed herein may be wet or dry compositions, and components may be incorporated into the food composition and / or incorporated on the surface of any composition component by, for example, spraying, agglomerating, sprinkling, or precipitation onto the surface. Furthermore, the dog food compositions may be formulated and manufactured to be of various forms and / or consistency. For example, the dog food composition may be, for example, a dry, moist, or semi-moist animal food composition. "Semi-moist" refers to a food composition containing about 25 to about 35% moisture. "Moist" food refers to a food composition having a moisture content of about 60 to 90% or more. "Dry" food refers to a food composition having a moisture content of about 3 to about 12% and is often manufactured in the form of pieces or kibble.

[0162] Foods may also contain multiple cohesive components, such as soft, chewy, meat-like particles or fragments, and kibble having an outer coating and an inner “core” component. In some embodiments, the dog food composition may be in the form of kibble or food kibble. As used herein, the terms “kibble” or “food kibble” refer to granular pellets of dog feed, such as components of dog feed, or dog feed. In some embodiments, food kibble has a moisture or water content of less than 15% by weight. Food kibble can have a wide range of textures, from hard to soft. Food kibble can have a wide range of internal structures, from puffy to dense. Food kibble may be formed by an extrusion or baking process. In non-limiting examples, food kibble may have a uniform or variable internal structure. For example, food kibble may include a core and coating to form coated kibble. It should be understood that when the terms "kibble" or "food kibble" are used, they can refer to either uncoated or coated kibble.

[0163] According to one aspect of the present disclosure, the dog food composition is in a form comprising a core and a coating placed on the core. In one aspect, the core comprises an alpha-amino acid (e.g., lysine) and one or more ancient grains (e.g., quinoa and amaranth), while the coating comprises an eggshell membrane.

[0164] Dog food compositions may include, for example, binders for maintaining kibble shape and / or binding a coating to the core. In certain embodiments, binders may include monosaccharides such as glucose, fructose, mannose, and arabinose; disaccharides and trisaccharides such as sucrose, lactose, maltose, trehalose, and lactulose; corn and rice syrup solids; dextrins such as corn, wheat, rice, and tapioca dextrin; maltodextrin; starches such as rice, wheat, corn, potato, tapioca starch, or these starches modified by chemical modification; alginates, chitosan; gums such as carrageenan and gum arabic; glycerol, sorbitol, mandarin Examples of polyols such as nitrol, xylitol, and erythritol; esters of polyols such as sucrose esters, polyglycol esters, glycerol esters, polyglycerol esters, and sorbitan esters; sorbitol; molasses; honey; gelatin; peptides; whey liquid, whey powder, concentrated whey, whey isolate, whey protein isolate, high-lactose whey by-products; meat broth solids such as chicken broth and chicken broth solids; soy protein; and proteins and denatured proteins such as egg white, or any combination thereof. In one embodiment, the dog food composition is substantially free of or does not contain egg white and / or whole egg components.

[0165] In certain embodiments, the binder includes, but is not limited to, lipids and / or lipid derivatives. Lipids can be used in combination with water and / or other binder components. Examples of lipids include vegetable fats such as soybean oil, corn oil, rapeseed oil, olive oil, safflower oil, palm oil, coconut oil, palm kernel oil and partially and fully hydrogenated derivatives thereof, animal fats and partially and fully hydrogenated derivatives thereof, and waxes.

[0166] The compositions of this disclosure may further contain other additives in amounts and combinations well known to those skilled in the art. Such additives should be present in amounts that do not impair the purpose and effects provided by the present invention. Examples of additives include substances that have a stabilizing effect, sensory stimulants, processing aids, and substances that provide nutritional benefits.

[0167] Examples of stabilizing substances include those that tend to increase the shelf life of the composition. Other examples of such additives that are potentially suitable for inclusion in the composition of the present invention include, for example, preservatives, antioxidants, cohesive agents and scavengers, packaging gases, stabilizers, emulsifiers, thickeners, gelling agents, and wetting agents. Examples of emulsifiers and / or thickeners include gelatin, cellulose ethers, starch, starch esters, starch ethers, and modified starch. Additives for coloring, flavoring, and nutritional purposes include colorants, salts (including, but not limited to, sodium chloride, potassium citrate, potassium chloride, and other edible salts), vitamins, minerals, and flavorings. Other additives include glucosamine, chondroitin sulfate, plant extracts, and herbal extracts.

[0168] The concentration of such additives in the composition can typically be up to about 5% by weight, based on the total weight of the dog food composition on a dry matter basis. In some embodiments, the concentration of such additives (especially when such additives are primarily nutritional balancers, such as vitamins and minerals) is about 0% to about 2.0% by weight, based on the total weight of the dog food composition on a dry matter basis. In some embodiments, the concentration of such additives (again, especially when such additives are primarily nutritional balancers) is about 0% to about 1.0% by weight, based on the total weight of the dog food composition on a dry matter basis. While the aforementioned list of additives may be potentially appropriate in some embodiments, one or more of the aforementioned additives may be excluded from other embodiments of the dog food composition.

[0169] Dog food compositions may be manufactured by various methods to achieve a desired dog food composition or desired form. For example, dry food may be baked or extruded and then cut into individual shapes such as kibble. In some embodiments, dog food compositions may be prepared in canned or wet form using conventional food preparation processes known to those skilled in the art. Typically, ground animal protein tissue is mixed with grains, a suitable carbohydrate source, fats, oils, and other ingredients such as balance ingredients (including special-purpose additives, e.g., vitamin and mineral mixtures, inorganic salts, cellulose, beet pulp, etc.) and a sufficient amount of water for processing. These ingredients are mixed in a container suitable for heating during the blending of ingredients. Heating of the mixture is carried out using any suitable method, such as direct steam injection or the use of a container equipped with a heat exchanger. After the addition of all ingredients of the composition, the mixture may be heated to a temperature of 50°F to 212°F. Temperatures outside this range may be used, but may not be commercially practical without other processing aids. When heated to a suitable temperature, the material is typically in the form of a viscous liquid and is dispensed into cans. The lids are attached and the containers are sealed. The sealed cans are then placed in a conventional apparatus designed to sterilize the contents. Sterilization is typically achieved by heating to a temperature above 230°C for an appropriate amount of time, depending on the temperature used, the properties of the composition, and related factors. The dog food compositions and foods of this disclosure may also be added to or combined with food compositions before, during, or after their preparation.

[0170] In some embodiments, the food may be prepared in a dry form using conventional processes known to those skilled in the art. Typically, dry components, including dried animal protein, vegetable protein, grains, etc., are ground and mixed together. Liquid or wet components, including fats, oils, water, animal protein, water, etc., are added and mixed with the dry material. Specific formulations, addition sequences, combinations, methods, and equipment used to combine various components can be selected from those known in the art. For example, in certain embodiments, the resulting mixture is processed into kibble or similar dry pieces, formed using an extrusion process, in which the mixture of dry and wet components is subjected to mechanical work at high pressure and temperature, passed through a small opening or aperture, and cut into kibble with a rotating knife, for example. The resulting kibble is dried and optionally coated with one or more topical coatings, including, for example, flavorings, fats, oils, powder components. Kibble can also be made from dough using a baking method instead of extrusion, in which the dough is placed in a mold and then subjected to a drying and heating process.

[0171] When preparing a composition, any component can generally be incorporated into the composition during the formulation process, for example, while and / or after mixing the other components of the composition. The distribution of these components into the composition can be achieved by conventional means. In certain embodiments, ground animal and / or poultry protein tissue is mixed with other components, including nutritional balancers and inorganic salts, and further mixed with other components, including cellulose, beet pulp, fillers, etc., with enough water for processing. [Examples]

[0172] The following embodiments are provided to further illustrate certain preferred embodiments of the present invention and should not be construed as limiting the scope of the invention.

[0173] Example 1 Fecal samples were collected from 24 dogs of varying ages, metabolic rates, sexes, body weights, fecal score GI phenotypes, and health conditions at the time of sample collection. See Table 1A below. [Table 1A] TIFF2026512800000003.tif52160

[0174] Data from 24 fecal samples, including microbiome data at three levels (genus (g), family (f), phylum (p)), SCFA, pH, and related metadata (a total of 96 variables), were included in PCA analysis. A segregation was observed between the "GI-risk" group and the more overlapping "GI-risk-free" group (Figures 1A and 1B). Specifically, components 1 and 2 explained only 22.9% and 20.2% of the variability, respectively. Evaluation of the 96 variables between "GI-risk" and "GI-risk-free" groups using PLSDA yielded 16 candidate biomarkers (see Table 2 below). Variables marked with an asterisk were positively associated with the "GI-risk-free" group, while variables marked with a number were negatively associated. Items in bold from Total SCFA to f_Enterobacteria seaae indicate the importance of the marked variables as distinguishing factors between the "GI-risk-free" and "GI-risk" groups based on VIP scores. Model quality: R2 cumulative: 0.537 and Q2 cumulative: 0.472. In Figure 1B, the numbering replaces the following labels. [Table 1B] [Table 2]

[0175] This panel of 16 biomarkers distinguished dogs at "GI risk" from those at "GI risk" with an AUC of 0.929 and a classification precision of 0.83 (Figure 2). In particular, the relative abundance of fifteen variables (total SCFA, C2, C3, C4, pH, moisture, fecal score, Philam bacteroidetes excluding F3:f_bacteroideae and F4:f_prevotellaceae, Prevotellaceae, Ruminococceae, Enterobacteriaceae, Bacteroideae, Prevotella, Shannon diversity, and equality) differed significantly between dogs at "GI risk" and those at "GI risk" (based on the Wilcoxon / Kruskas-Wallis test, p<0.05) (Figure 3).

[0176] Furthermore, this model exhibits a sensitivity of 0.916 and a specificity of 0.75 for detecting individuals with "GI risk" and "GI risk-free" states, respectively. PCA was also performed using 16 biomarkers identified by the PLSDA model, demonstrating good segregation between the "GI risk" and "GI risk-free" populations (Figures 4A and 4B). Of these 16 biomarkers, the variable "pH" was negatively associated with disease status ("GI risk" vs. "GI risk-free"). Correlation analyses of other biomarkers compared to "pH" showed that all other biomarkers were significantly correlated with "pH," either positively or negatively (Figures 5A and 5B, and Table 3). [Table 3]

[0177] Example 2: Collection of fecal samples A fecal score of 5 indicates good (firm), and a fecal score of 1 indicates poor (watery). Five fecal samples were collected from each individual dog for each fecal score grade from 1 to 5. Fecal samples were collected immediately after defecation (within 30 minutes), and all feces were transferred to a labeled collection bag for homogenization. The samples were completely homogenized, dispensed into labeled cryovials, and flash-frozen in liquid nitrogen. The flash-frozen tubes were then kept at -70°C until further processing. 0Moved to C. The sample numbers and associated metadata are listed in Table 2.

[0178] Example 3: Measurement of moisture, pH, and short-chain fatty acids (SCFAs) The pH of feces was measured using a digital pH meter (Hach Model H160 pH meter) equipped with a pH probe (ISFET Series H Metal Probe) designed for measuring pH within meat. The pH meter was calibrated with three sample sets (pH 4.0, pH 7.0, and pH 10.0).

[0179] SCFAs in feces were measured using gas chromatography with liquid-liquid extraction and flame ionization detection to quantify specific carboxylic acids in feces. Briefly, fecal samples were measured in a centrifuge tube with an internal standard solution and then extracted with aqueous buffer. Aliquots of the extract were acidified and then extracted with an organic solvent. The organic extract was separated from the aqueous phase and analyzed using gas chromatography. LAB-RES-082.0: pH of stool LAB-RES-45.2: Measurement of volatile carboxylic acids in feces

[0180] Example 4: 16S rRNA amplicon sequencing Total fecal DNA was extracted from frozen fecal samples using the MoBio PowerFecal DNA Extraction Kit. Following total DNA extraction, 16S rDNA amplicons were constructed from the samples using PCR with primer sets spanning the V3 and V4 hypervariable regions, and then qualitatively analyzed using an Agilent 2100 Bioanalyzer. Index PCR was performed, followed by library quantification, normalization, and sample pooling according to the manufacturer's instructions. The final libraries of pooled samples were loaded into MiSeq v3 sample-filled cartridge kits, and the cartridges were placed in a MiSeq (Illumina) sequencer for sequencing the amplicons. Sample sequences were demultiplexed using the "metagenomics" workflow on the MiSeq to obtain FASTQ files. The FASTQ files were processed using QIIME software to classify sequence reads using the Greengenes database and to create genus abundances.

[0181] Total DNA was extracted from frozen fecal samples using the PowerFecal DNA Isolation Kit (MoBio, Carlsbad, California) according to the modified manufacturer's instructions, which included introducing a sonication step followed by vortexing the bead tube containing the fecal sample horizontally for 15 minutes. PCR amplification was performed in a Biorad C1000 Touch thermal cycler under the following conditions: 25 cycles, 30 seconds at 95°C, 30 seconds at 55°C, and 45 seconds at 72°C, followed by 5 minutes at 72°C, using primer pairs 347F and 803R spanning the V3-V4 hypervariable region of the 16s rRNA gene with an Illumina adapter (Nossa et al 2013). The amplicons were purified using Agencourt AmPure XP beads (Beckman Coulter) according to the manufacturer's instructions, and their concentrations were determined using a Qubit Fluorescence Spectrometer 3.0 (Life Technologies) with a high-sensitivity DNA kit. Furthermore, amplicon quality was determined using an Agilent 2100 bioanalyzer. Index PCR, library quantification, normalization, and pooling were performed, followed by Illumina's 16S metagenomics sequencing library preparation protocol (part number 15044223 Rev.A). The libraries were mixed with an Illumina-prepared PhiX control library (final concentration 10%) and denatured using fresh NaOH. The pooled final denatured libraries (6 pm) were loaded into a Miseq v3 reagent cartridge kit and paired-end sequencing was performed for 2 / 251 cycles using Miseq Control Software (MCS) 2.4, RTA 1.18.54, and Miseq Reporter 2.4. For each Miseq run, one simulated community sample was included as a positive control, and water was used as a negative control. Both positive and negative control samples were processed identically to fecal DNA samples for the full sequencing protocol. LAB-LS-060.1: DNA isolation from bacteria in dog / cat feces - MoBio PowerFecal kit.

[0182] Example 5: Data Analysis The Shannon diversity (H) index was calculated, including all identified operational taxa (OTUs) in the sample. Those with ubiquity below 75% were excluded from the data analysis. For the remaining 46 genera (Table 4), zero counts were imputed, and then all counts were converted to relative abundance by dividing each individual count by the total number of counts in the sample. The relative abundances were converted to centered log-reverse ratios (CLR) for further statistical analysis. A similar approach was followed for phylum and family-level microbiome data. The R package, ALDEx2, was used for imputation and conversion.

[0183] Partial least squares discriminant analysis (PLS-DA) was performed to classify animals as either "GI risk" or "GI risk-free" using microbiome CLR, SCFA, pH, and associated metadata. XLSTAT was used for the PLS-DA analysis. [Table 4]

[0184] While the present invention has been described in relation to specific embodiments, including currently preferred modes for carrying out the invention, those skilled in the art will recognize that there are numerous variations and rearrangements of the methods and systems described above.

[0185] It should be understood that other embodiments may be utilized and that structural and functional modifications may be made without departing from the scope of the present invention. Therefore, the spirit and scope of the present invention should be interpreted broadly as set forth in the appended claims.

Claims

1. A method for identifying whether a target dog is at risk of or not at risk of gastrointestinal (GI) disease, (a) Analyze fecal samples from the subject dogs to determine the values ​​of the fecal samples for one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viiii) the abundance of bacteria Prevotellaceae, Ruminococcusceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes. (b) Compare the values ​​obtained from (a) with the values ​​obtained from one or more dogs without risk of GI disease for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture, (vii) fecal score, and (viiii) bacterial abundance of Prevotellaceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes. A method comprising the above, wherein if one or more of the values ​​in the fecal sample of the subject dog are similar to or identical to the control value, there is no risk of GI disease.

2. The analysis described above (a) is to determine the values ​​of the fecal sample for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viiii) the abundance of Prevotella seaae, Ruminococca seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes. The method according to claim 1, wherein (b) the comparison is made with respect to a control value for one of the following: (i) total short-chain fatty acid content (SCFA), (ii) level of C2 SCFA, (iii) level of C3 SCFA, (iv) level of C4 SCFA, (v) pH, (vi) moisture, (vii) fecal score, and (viiii) bacterial abundance of Prevotella seaae, Ruminococcus seaae, Bacteroida seaae, Enterobacteria seaae, and Bacteroidetes.

3. The method according to claim 2, wherein if each of the values ​​of the fecal sample of the subject dog is similar to or identical to the control value, there is no risk of GI disease.

4. The method according to claim 2, wherein if each of the values ​​of the fecal sample of the subject dog differs from the control value, the subject dog is at risk of GI disease.

5. The subject dog, when the fecal sample was compared to the control value, (i) Total content of short-chain fatty acids at approximately equivalent or higher levels, (ii) C2 SCFA at a nearly equivalent or higher level, (iii) C3 SCFA at a nearly equivalent or higher level, (iv) C4 SCFA at approximately the same or higher level, (v) Approximately the same or lower pH, (vi) a fecal score that is nearly equivalent or higher, and (vii) The method according to claim 2, wherein there is no risk of GI disease if there is a substantially equal or higher abundance of Prevoteraceae, Ruminococcaceae, and Bacteroidaceae bacteria, and a substantially equal or lower abundance of Enterobacteriaceae and Bacteroidetes bacteria other than Prevoteraceae and Bacteroidaceae.

6. The subject dog, when the fecal sample was compared to the control value, (i) Total content of short-chain fatty acids at a lower level, C2 SCFA, a lower level than (ii) C3 SCFA, a lower level than (iii). C4 SCFA at a lower level than (iv), (v) higher pH, (vi) Lower fecal score than, and The method according to claim 2, wherein there is a risk of GI disease if there are Prevoteraceae, Ruminococcaceae, and Bacteroidaceae at lower abundances than (vii), and Enterobacteriaceae and Bacteroidetes other than Prevoteraceae and Bacteroidaceae at higher abundances.

7. The subject dog, when the fecal sample is compared to the control value, has at least one of the following characteristics: (i) Total content of short-chain fatty acids at approximately equivalent or higher levels, (ii) C2 SCFA at a nearly equivalent or higher level, (iii) C3 SCFA at a nearly equivalent or higher level, (iv) C4 SCFA at approximately the same or higher level, (v) Approximately the same or lower pH, (vi) A fecal score that is nearly the same or higher, (vii) The method according to claim 1, wherein there is no risk of GI disease if there is a substantially equal or higher abundance of Prevoteraceae, Ruminococcaceae, and Bacteroidaceae bacteria, or a substantially equal or lower abundance of Enterobacteriaceae other than Prevoteraceae and Bacteroidaceae and Bacteroidetes bacteria.

8. The subject dog, when the fecal sample is compared to the control value, has at least one of the following characteristics: (i) Total content of short-chain fatty acids at a lower level, C2 SCFA, a lower level than (ii) C3 SCFA, a lower level than (iii). C4 SCFA at a lower level than (iv), (v) higher pH, A fecal score lower than (vi), or The method according to claim 1, wherein there is a risk of GI disease if the abundance of bacteria Prevotellaceae, Ruminococcaceae, and Bacteroidaceae is lower than (vii), or if the abundance of bacteria Enterobacteriaceae other than Prevotellaceae and Bacteroidaceae and Bacteroidetes is higher.

9. The method according to claim 1, wherein the fecal sample is analyzed for the abundance of bacteria Prevoteraceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes, and the amplification of species-specific 16S rDNA within each of Prevoteraceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes.

10. A kit for identifying whether a target dog is at risk of or not of gastrointestinal (GI) disease by obtaining the results of fecal analysis, wherein the kit comprises at least one of the following: a reagent or apparatus for determining the pH of a fecal sample; one or more reagents or apparatus for measuring volatile carboxylic acids in a fecal sample; and one or more reagents or apparatus for determining the abundance of one or more bacteria in a fecal sample from Prevotellaseae, Luminococcusceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes, and the results are obtained by performing the above.

11. The kit according to claim 10, comprising the reagent or apparatus for determining the pH of a fecal sample, one or more reagents or apparatus for measuring volatile carboxylic acids in a fecal sample, and each of the one or more reagents or apparatus for determining the abundance of each of the bacteria Prevotellaceae, Luminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes in a fecal sample.

12. The kit according to claim 10, comprising one or more reagents for determining the abundance of Enterobacteria seae and Prevotella seae in the sample.

13. The kit according to claim 10, wherein the one or more reagents or apparatus for determining the abundance of one or more of Prevoteraceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes in a fecal sample comprises PCR primers for amplifying 16S rDNA from the target bacteria in Prevoteraceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes.

14. The device further comprises inputs, memory, a processor, and outputs, The input is configured to receive the result, The memory includes processor-executable instructions for comparing the result with a control value, and for preparing a conclusion that there is or is not a risk based on the comparison, and for outputting the result. The processor is configured to execute the processor-executable instructions, The kit according to any one of claims 10 to 13, wherein the output is configured to display the conclusion.

15. The kit according to claim 14, wherein the processor-executable instructions further include instructions that display a recommendation to feed the subject dog a GI disease mitigation diet when it is concluded that the subject dog is at risk, and the output is further configured to display the recommendation.

16. A method for preventing or treating a GI disease, comprising: identifying whether a subject dog is at risk of or not of a gastrointestinal (GI) disease; and, if the identifying step concludes that the subject dog is at risk, providing the subject dog with a GI disease relief diet.

17. The step of identifying whether the target dogs are at risk or not of gastrointestinal (GI) disease is: (a) Analyze fecal samples from the subject dogs to determine the values ​​of the fecal samples for one or more of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture content, (vii) fecal score, and (viiii) the abundance of bacteria Prevotellaceae, Ruminococcusceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes. (b) Comparing the values ​​obtained from (a) to control values ​​from one or more dogs without risk of GI disease for each of the following: (i) total short-chain fatty acid content (SCFA), (ii) C2 SCFA level, (iii) C3 SCFA level, (iv) C4 SCFA level, (v) pH, (vi) moisture, (vii) fecal score, and (viiii) bacterial abundance of Prevotellaceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes, The method according to claim 16, wherein if one or more of the values ​​in the fecal sample of the subject dog are similar to or the same as the control value, there is no risk of GI disease, or if one or more of the values ​​in the fecal sample of the subject dog are different from the control value, there is a risk of GI disease.

18. The method according to claim 17, wherein if the values ​​of each of the fecal samples of the subject dog are similar to or the same as the control values, there is no risk of GI disease, or if the values ​​of each of the fecal samples of the subject dog are different from the control values, there is a risk of GI disease.

19. The subject dog, when the fecal sample was compared to the control value, (i) Total content of short-chain fatty acids at approximately equivalent or higher levels, (ii) C2 SCFA at a nearly equivalent or higher level, (iii) C3 SCFA at a nearly equivalent or higher level, (iv) C4 SCFA at approximately the same or higher level, (v) Approximately the same or lower pH, (vi) a fecal score that is nearly equivalent or higher, and (vii) The method according to claim 17, wherein there is no risk of GI disease if there is a substantially equal or higher abundance of Prevoteraceae, Ruminococcaceae, and Bacteroidaceae bacteria, and a substantially equal or lower abundance of Enterobacteriaceae other than Prevoteraceae and Bacteroidaceae and Bacteroidetes bacteria.

20. The subject dog, when the fecal sample was compared to the control value, (i) Total content of short-chain fatty acids at a lower level, C2 SCFA, a lower level than (ii) C3 SCFA, a lower level than (iii). C4 SCFA at a lower level than (iv), (v) higher pH, (vi) Lower fecal score than, and The method according to claim 17, wherein there is a risk of GI disease if the bacterial abundance of Prevotellaceae, Ruminococcaceae, and Bacteroidaceae is lower than (vii), and the bacterial abundance of Enterobacteriaceae and Bacteroidetes other than Prevotellaceae and Bacteroidaceae is approximately higher.

21. The subject dog, when the fecal sample is compared to the control value, has at least one of the following characteristics: (i) Total content of short-chain fatty acids at approximately equivalent or higher levels, (ii) C2 SCFA at a nearly equivalent or higher level, (iii) C3 SCFA at a nearly equivalent or higher level, (iv) C4 SCFA at approximately the same or higher level, (v) Approximately the same or lower pH, (vi) A fecal score that is nearly the same or higher, (vii) The method according to claim 17, wherein there is no risk of GI disease if there is a substantially equal or higher abundance of Prevoteraceae, Ruminococcaceae, and Bacteroidaceae bacteria, and a substantially equal or lower abundance of Enterobacteriaceae other than Prevoteraceae and Bacteroidaceae and Bacteroidetes bacteria.

22. The subject dog, when the fecal sample is compared to the control value, has at least one of the following characteristics: (i) Total content of short-chain fatty acids at a lower level, C2 SCFA, a lower level than (ii) C3 SCFA, a lower level than (iii). C4 SCFA at a lower level than (iv), (v) higher pH, A fecal score lower than (vi), or The method according to claim 17, wherein there is a risk of GI disease if the bacterial abundance of Prevotellaceae, Ruminococcaceae, and Bacteroidaceae is lower than (vii), and the bacterial abundance of Enterobacteriaceae and Bacteroidetes other than Prevotellaceae and Bacteroidaceae is approximately higher.

23. The method according to claim 17, wherein the fecal sample is analyzed for the abundance of bacteria Prevoteraceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes, and the amplification of species-specific 16S rDNA in each of Prevoteraceae, Ruminococcaceae, Bacteroidaceae, Enterobacteriaceae, and Bacteroidetes.

24. The aforementioned diet for alleviating GI disease is Approximately 1% to 10% by weight of quinoa, Approximately 1% to 10% by weight of amaranth, Approximately 0.5% to 8% by weight of eggshell membrane, and The dog food composition comprises approximately 2% to approximately 20% by weight of dietary fiber, wherein the dietary fiber is i) Approximately 1% to 18% by weight of insoluble fibers, and ii) Contains approximately 0.5% to 8% by weight of soluble fibers, The method according to any one of claims 16 to 23, wherein all weight percentages are based on the total weight of the dog food composition on a dry matter basis.

25. The method according to claim 24, wherein the dog food composition further comprises about 0.1% to about 10% by weight of fatty acids.

26. The method according to claim 25, wherein the fatty acid includes a polyunsaturated fatty acid.

27. The method according to claim 26, wherein the polyunsaturated fatty acid comprises an omega-3 fatty acid, an omega-6 fatty acid, or a combination of two or more thereof.

28. The method according to claim 26, wherein the polyunsaturated fatty acid comprises an omega-3 fatty acid selected from linolenic acid, stearidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid, or docosahexaenoic acid, or a combination of two or more thereof.

29. The method according to claim 27 or 28, wherein the omega-3 fatty acid comprises alpha-linolenic acid.

30. The method according to claim 27 or 28, wherein the omega-3 fatty acid comprises eicosapentaenoic acid, docosahexaenoic acid, or a combination thereof.

31. The method according to claim 26, wherein the polyunsaturated fatty acid comprises an omega-6 fatty acid selected from linolenic acid, calendic acid, eicosadienoic acid, arachidonic acid, docosadienoic acid, adrenaline, osbondic acid, tetracosatetraenoic acid, or tetracosapentaenoic acid, or a combination of two or more thereof.

32. The method according to claim 26, wherein the polyunsaturated fatty acid comprises an omega-6 fatty acid selected from linolenic acid, arachidonic acid, and two or more combinations thereof.

33. A device comprising an input, memory, processor, and output, The input is configured to receive the results of a method for identifying whether the target dog is at risk or not of gastrointestinal (GI) disease. The memory includes processor-executable instructions for comparing the result with a control value, and for preparing a conclusion that there is or is not a risk based on the comparison, and for outputting the result. The processor is configured to execute the processor-executable instructions, A device wherein the output is configured to display the conclusion.

34. The apparatus according to claim 33, wherein the processor-executable instructions further include instructions for displaying a recommendation to feed the subject a GI disease relief diet when it is concluded that the subject dog is at risk, the processor is further configured to execute instructions for displaying a recommendation, and the output is further configured to display the recommendation.

35. The aforementioned recommendation for feeding the aforementioned subjects a diet to alleviate GI disease is, Approximately 1% to 10% by weight of quinoa, Approximately 1% to 10% by weight of amaranth, Approximately 0.5% to 8% by weight of eggshell membrane, and This is for feeding a dog food composition containing approximately 2% to 20% by weight of dietary fiber, wherein the dietary fiber is i) Approximately 1% to 18% by weight of insoluble fibers, and ii) Contains approximately 0.5% to 8% by weight of soluble fibers, The apparatus according to claim 34, wherein all weight percentages are based on the total weight of the dog food composition on a dry matter basis.

36. The apparatus according to claim 34, wherein the recommendation for feeding the subject a GI disease relief diet is for feeding the subject a dog food composition comprising the dog food composition described in any one of claims 24 to 32.

37. The apparatus according to any one of claims 33 to 36, wherein the method includes the method according to any one of claims 1 to 13.

Images