# Patent

## Title
Methods for inhibiting a decline in learning and/or memory in animals

## Abstract

A method for inhibiting a decline in learning and/or memory in an animal comprising maintaining the animal on an antioxidant-fortified diet for a period of at least about 3 years. The diet comprises at least one antioxidant in a total antioxidant amount effective to achieve such inhibition. Suitable antioxidants include vitamin E, vitamin C, α-lipoic acid and antioxidant-containing plant meals. The methods are particularly useful for young adult canines such as dogs.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims priority to U.S. Provisional Application Ser. No. 60/639,928, filed Dec. 22, 2004, and to U.S. Provisional Application Ser. No. 60/669,097, filed Apr. 7, 2005, the disclosures of which is incorporated herein by reference.


BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates generally to methods for inhibiting a decline in learning and/or memory in animals and particularly to methods for using antioxidants inhibiting a decline in learning and/or memory in canine and feline animals.
2. Description of the Related Art
In designing foods for animals such as dogs and cats, optimal physical health through good nutrition is an important goal. However, a diet which meets ordinary physical nutritional requirements may not be enough to provide the best nutrition for maximizing cognitive function, including learning and memory.
Dogs, cats, and many other animals are required to utilize their cognitive skills for many purposes in their interactions with humans, other animals and/or their environment. For example, working dogs (such as police dogs and guard dogs), hunting dogs, herding dogs (including sheepdogs), guide dogs, show dogs and search and rescue dogs are expected to demonstrate even higher levels of cognitive skills than dogs kept only as pets. Dogs and other animals of any age can demonstrate declines in learning ability and memory, resulting in frustration on the part of the owner or caregiver of the animal and the animal.
Decline in memory and learning ability in animals has largely been accepted as part of the cost of working and living with animals. Such declines are generally age-related and become most noticeable in aged and geriatric animals, but can also be an important limitation to the usefulness of working animals such as dogs and the enjoyment and companionship provided by pets and other in their young adult years, for example from about 1 to about 6 years of age. Given the problem, there is a need for new methods for inhibiting a decline in learning and/or memory in animals such as dogs and cats.
SUMMARY OF THE INVENTION
The invention provides methods for inhibiting a decline in learning and/or memory in animals, particularly canines and felines such as dogs and cats, by maintaining the animal on an antioxidant-fortified diet comprising a decline in learning and/or memory inhibiting amount of at least one antioxidant for a period of at least about 3 years. The methods are useful for animals such as canines and felines of any breed of occupation but are particularly useful for dogs and cats that rely on maintenance of cognitive functions such as learning and memory during their adult years for effectiveness or quality of companionship or for providing useful services to their owner or caregiver.
Additional or alternative advantages and benefits of the present method will be apparent to one skilled in the art from reading this specification.


DETAILED DESCRIPTION OF THE INVENTION
Definitions
The term “animal” means any animal susceptible to or suffering from a decline in learning and/or memory, particularly resulting from aging.
A “young adult” animal for a canine or feline means a canine or feline of about 1 to about 6 years of age. For other animal species, the age range for a “young adult” will vary according to the species, but will be known to skilled artisans.
The phrase “total antioxidant amount” stated herein as being effective for a given function, e.g., to inhibit decline in learning and/or memory, refers to an amount in total of all antioxidants, or according to the context all specified antioxidants, present in or added to the diet. If only one antioxidant is present, the “total antioxidant amount” is the amount of that one antioxidant.
The term “single package” means that the components of a kit are physically associated in or with one or more containers and considered a unit for manufacture, distribution, sale, or use. Containers include, but are not limited to, bags, boxes, bottles, shrink wrap packages, stapled or otherwise affixed components, or combinations thereof. A single package may be containers of individual antioxidants physically associated such that they are considered a unit for manufacture, distribution, sale, or use.
The term “virtual package” means that the components of a kit are associated by directions on one or more physical or virtual kit components instructing the user how to obtain the other components, e.g., in a bag containing one component and directions instructing the user to go to a website, contact a recorded message, view a visual message, or contact a caregiver or instructor to obtain instructions on how to use the kit.
The Invention
The invention provides methods for inhibiting a decline in learning and/or memory in animals. The methods comprise maintaining the animal on an antioxidant-fortified diet comprising a decline in learning and/or memory inhibiting amount of at least one antioxidant for a period of at least about 3 years. The methods surprisingly inhibit a decline in learning and/or memory in animals and increase their mental capacity.
The methods of the invention are useful for a variety of human and non-human animals, including avian, bovine, canine, equine, feline, hicrine, murine, ovine, and porcine animals, and are particularly useful for companion animals such as canines and felines, including dogs and cats.
In one embodiment, the animal is a canine and the diet comprises at least one antioxidant selected from the group consisting of vitamin E, vitamin C, α-lipoic acid, and combinations thereof. In another, the animal is a feline and the diet comprises at least one antioxidant selected from the group consisting of vitamin E, vitamin C, and combinations thereof. In either embodiment, the diet may further comprise plant meal such as fruit meal and/or vegetable meal.
The methods are especially useful for dogs maintained on the fortified diet for an extended period. The benefit will become greater the longer the dog is maintained on the fortified diet. For example, maintaining the dog on such a diet for a period of at least about 4 years or a period of at least about 5 years or for substantially the duration of the dog's adult life can further enhance the benefits obtained.
In some embodiments, a single antioxidant is present in the diet. In others, a plurality of antioxidants is present in the diet.
Antioxidants useful in the present invention include any substance that is capable of reacting with free radicals and neutralizing them. Illustrative examples of such substances include β-carotene, selenium, coenzyme Q10 (ubiquinone), luetin, tocotrienols, soy isoflavones, S-adenosylmethionine, glutathione, taurine, N-acetylcysteine, vitamin E, vitamin C, α-lipoic acid and L-carnitine. Examples of foods containing useful levels of one or more antioxidants include but are not limited to ginkgo biloba, green tea, broccoli, citrus pulp, grape pomace, tomato pomace, carrot, spinach, and a wide variety of fruit meals and vegetable meals.
Except where the context demands otherwise, the term “vitamin E” is used generically herein to encompass any tocopherol or tocotriene compound, including any enantiomer or racemate thereof, and any mixture of such compounds, having vitamin E activity, including α-tocopherol ((+)-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-6-chromanol), β-tocopherol ((+)-2,5,8-trimethyl-2-(4,8,12-trimethyltridecyl)-6-chromanol), γ-tocopherol ((+)-2,7,8-trimethyl-2-(4,8,12-trimethyltridecyl)-6-chromanol), δ-tocopherol ((+)-8-methyl-2-(4,8,12-trimethyltridecyl)-6-chromanol), α-tocotrienol (2,5,7,8-tetramethyl-2-(4,8,12-trimethyl-3,7,11-tridecatrienyl)-6-chromanol) and β-tocotrienol (2,5,8-trimethyl-2-(4,8,12-trimethyl-3,7,11-tridecatrienyl)-6-chromanol). Vitamin E can be administered as any one or a mixture of the above compounds or in the form of various derivatives thereof such as esters, including vitamin E acetate, succinate, palmitate and the like, that exhibit vitamin E activity after ingestion by the animal. Typically vitamin E as used in the present method comprises α-tocopherol or an ester thereof.
Vitamin C can be administered as ascorbic acid, e.g., L-ascorbic acid, or as various derivatives thereof such as calcium phosphate salt, cholesteryl salt, and ascorbate-2-monophosphate. Salts of vitamin C include, for example, sodium salt, calcium salt, zinc salt and ferrous salt. Esters include, for example, stearate, palmitate and like derivatives. Vitamin C or a derivative thereof can be in any physical form, for example, a liquid, a semisolid, a solid, or a heat stable form that exhibits vitamin C activity after ingestion by the animal.
Lipoic acid can be administered as such, as a lipoate salt or ester, or as a lipoate derivative, for example as described in U.S. Pat. No. 5,621,117. As used herein, “lipoic acid” is synonymous with α-lipoic acid and can be provided in various forms including racemic mixtures, salts, esters and/or amides thereof.
Antioxidants can be added to the diet as antioxidant substances per se (including derivatives thereof having antioxidant activity), such substances being of natural, biosynthetic or chemosynthetic origin. Alternatively or in addition, antioxidants can be added as components of food ingredients such as those listed above.
In some embodiments, including animals with life spans greater than about 15 years, the animal is maintained on the antioxidant-fortified diet for a period of greater than 3 years, e.g., for a period of at least about 4 years or a period of at least about 5 years.
In one embodiment, the method comprises maintaining a dog on an antioxidant-fortified diet for a period of at least about 3 years, wherein the diet comprises at least one antioxidant and further comprises fruit and/or vegetable meal.
Diets include food compositions such as canned moist foods, extruded dry foods, supplements and treats, having the antioxidant as an ingredient thereof. Any food component of the diet can benefit from inclusion of an antioxidant as defined herein, but in one embodiment the food is one having a major (at least about 25%, for example at least about 50%, by weight) component derived from animal (e.g., mammal, bird, fish or seafood) proteinaceous tissues including muscle tissues and/or offal, optionally with a carbohydrate source such as cereal grains.
The antioxidant(s) such as vitamin E, vitamin C, α-lipoic acid and/or plant meal can be distributed more or less homogeneously through a food component of the diet. Alternatively, the antioxidant(s) can be present in a food component of the diet in whole or in part on surfaces of food pieces such as meat chunks, dry kibbles or individual treats such as biscuits.
The antioxidant(s) such as vitamin E, vitamin C, α-lipoic acid and/or plant meal should be present in an amount that is not toxic or otherwise deleterious to the health of an animal consuming a normal quantity of the food composition. In particular, the antioxidant(s) should be present at a concentration that does not cause undesirable effects on digestion, particularly long-term effects lasting several days or longer. Undesirable effects on digestion can include constipation or diarrhea.
An animal can be maintained on the antioxidant-fortified diet by feeding the antioxidant or mixture of antioxidants as a component of the animal's food or as a food supplement. The quantities provided in the food, all on a dry matter basis, are stated herein as the active material, i.e., measured as the free antioxidant substance. The antioxidant amount should not exceed a maximum above which toxicity is brought about. The antioxidant, or combination thereof, is fed to the animal in a total antioxidant amount effective to inhibit decline in learning and/or memory. What constitutes an effective amount varies depending on the breed and activity of the animal, the type of antioxidant(s) and other factors. One of skill in the art will, by routine testing based on the disclosure herein, readily establish a total antioxidant amount effective to inhibit decline in learning and/or memory in any particular situation.
Suitable amounts of vitamin E will normally be at least about 10 parts per million (ppm), more generally at least about 100 ppm, illustratively about 100 ppm to about 5000 ppm, about 150 ppm to about 2500 ppm, or about 200 ppm to about 1500 ppm, for example about 500 ppm to about 1000 ppm.
Suitable amounts of vitamin C will normally be at least about 5 ppm, more generally at least about 10 ppm, illustratively about 10 ppm to about 10,000 ppm, about 20 ppm to about 2000 ppm, or about 25 ppm to about 500 ppm, for example about 50 ppm to about 500 ppm or about 75 ppm to about 500 ppm.
Suitable amounts of α-lipoic acid will normally be at least about 5 ppm, more generally at least about 25 ppm, illustratively about 25 ppm to about 1000 ppm, or about 50 ppm to about 600 ppm, for example about 100 ppm to about 600 ppm.
Suitable amounts of plant meal will depend on its antioxidant content, determined for example as Trolox equivalent. Illustratively, a plant meal with an antioxidant content of at least about 25 μmol Trolox equivalents per gram of dry matter, or a combination of such plant meals, can be added to a diet at about 1% to about 5% by weight. Examples of plant meals having suitable antioxidant content include spinach pomace, tomato pomace, citrus pulp, grape pomace, carrot granules, broccoli, green tea, ginkgo biloba and corn gluten meals.
In one embodiment, the food further comprises one or more ingredients other than an antioxidant that, in combination with the antioxidant, can inhibit a decline in learning and/or memory in animals such as young adult dogs and cats. An example of such an ingredient is an omega-3 fatty acid such as eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA).
Where the antioxidant is localized on surfaces of food pieces, local concentrations can exceed those suggested here, but the overall concentration in the diet as a whole will generally be in a range as stated above.
In preparing a food useful according to the present invention, the components of the food are adjusted so that the antioxidant, for example vitamin E, vitamin C, α-lipoic acid and/or plant meal, is present in the food at a desired concentration. The antioxidant(s) can, for example, be incorporated into the food during formulation, such as during and/or after mixing of other components of the composition. Distribution of these components into the composition can be accomplished by any conventional method including standard mixing procedures.
Foods useful in the method of the invention can be prepared in a wet or containerized (e.g., canned or in pouches) form using conventional pet food processes. In one embodiment, ground animal (e.g., mammal, poultry, fish and/or seafood) proteinaceous tissues are mixed with other ingredients, including for example animal fats and vegetable oils, cereal grains, other nutritionally balancing ingredients, special purpose additives (e.g., vitamin and mineral mixtures, inorganic salts, cellulose and beet pulp, bulking agents, and the like); and water sufficient for processing is also added. These ingredients typically are mixed in a vessel suitable for heating while blending the components. Heating of the mixture can be effected in any suitable manner, such as, for example, by direct steam injection or by using a vessel fitted with a heat exchanger. Following addition of the last of these ingredients, the mixture is heated in a pre-cooking step to a temperature of up to about 100° C. Higher temperatures can be acceptable, but can be commercially impractical without use of other processing aids. When heated to the appropriate temperature, the material is typically in the form of a thick liquid. The thick liquid is filled into suitable containers such as cans, jars, pouches or the like. A lid is applied, and the container is hermetically sealed. The sealed containers are then placed into conventional equipment designed to sterilize the contents. This is usually accomplished by heating to a temperature of at least about 110° C. for an appropriate time, which is dependent on, for example, the temperature used and the composition. Products can also be prepared by an aseptic process wherein the contents are heated to commercial sterility before being packaged in sterilized containers.
The antioxidant(s) can be added to containerized food products before, during or after the pre-cooking step.
Foods useful in the method of the invention can be prepared in a dry form using conventional processes. In one embodiment, dry ingredients, including, for example, animal protein sources, plant protein sources, grains, etc., are ground and mixed together. Moist or liquid ingredients, including fats, oils, animal protein sources, minerals, water, etc., are then added to and mixed with the dry mix. The mixture is then processed into kibbles or similar dry pieces. Kibble is often formed using an extrusion process in which the mixture of dry and wet ingredients is subjected to mechanical work at a high pressure and temperature, and forced through small openings and cut off into kibble by a rotating knife. The wet kibble is then dried and optionally coated with one or more topical coatings which can include, for example, flavors, fats, oils, powders, and the like. Kibble also can be made from the dough using a baking process, rather than extrusion, wherein the dough is placed into a mold before dry-heat processing. Kibble also can be made from a food matrix undergoing pelletization.
The antioxidant(s) can be incorporated into the food by addition to the above-described mixtures before extrusion or by coating extruded kibble or pellets with the antioxidant(s) as an ingredient of a topical coating. For example, vitamin E can be added to liquids in a dry processing line, to a pre-conditioner composition or to a coating composition.
In another embodiment, a food useful according to the present method is a treat comprising at least one antioxidant, for example vitamin E, vitamin C, α-lipoic acid and/or plant meal. Treats include, for example, foods that are given to an animal to entice the animal to eat during a non-meal time. Treats for canines include biscuits, for example in the shape of dog bones. Treats can be nutritional, comprising one or more nutrients, and can, for example, comprise ingredients as described above for a food. Non-nutritional treats encompass any other treats that are non-toxic. The antioxidant(s) can be present in a coating on the surface of the treat, or incorporated into the treat, or both. Plant meal if present is typically incorporated into the treat.
In another embodiment, a food useful according to the present method is a nutritional supplement comprising at least one antioxidant, for example vitamin E, vitamin C, α-lipoic acid and/or plant meal, as defined herein. Supplements include, for example, a food used with another food to improve the nutritive balance or performance of the total. Supplements include foods that are fed undiluted as a supplement to other foods, offered free choice with other parts of an animal's diet that are separately available, or diluted and mixed with an animal's regular food to produce a complete diet. The AAFCO, for example, provides a discussion relating to supplements in the American Feed Control Officials, Inc. Official Publication (2003), at p. 220. Supplements can be in various forms including, for example, powders, liquids, syrups, pills, encapsulated compositions, etc.
Antioxidants such as vitamin E, vitamin C and α-lipoic acid can be encapsulated in food systems. For example, vitamin E can be added to a palatant (such as a digest or broth) at any time during processing of the palatant.
Antioxidants such as vitamin E, vitamin C, α-lipoic acid and/or plant meal can be added during manufacturing of the food, for example by a process of mixing or coating as indicated above. In one embodiment, the antioxidant(s) are added to an animal's food by the person responsible for feeding the animal. For this purpose, the antioxidant(s) can be added in diluted or dispersed form in a suitable carrier such as vegetable oil or edible powder as it is more convenient and reduces risk of accidental over-addition of the antioxidant. A powder comprising the antioxidant(s) can be sprinkled on an animal's food immediately before feeding. Alternatively, a liquid comprising the antioxidant(s) can be sprayed on the food. Such powder or liquid compositions are described herein as “antioxidant compositions” and can be thought of as “additives”. They can be applied to the top of a serving of food and/or can, if desired, be mixed into the food. The use of antioxidant compositions or additives, for example comprising vitamin E, vitamin C, α-lipoic acid and/or plant meal, as specified herein to inhibit decline in learning and/or memory in young adult dogs is an embodiment of the present invention. Accordingly there is provided herein a method for inhibiting a decline in learning and/or memory in a young adult dog, the method comprising adding to a food component of the diet of the dog an antioxidant composition that comprises a powder or liquid carrier having the antioxidant diluted or dispersed therein, and maintaining the dog on the diet for a period of at least about 3 years. A total antioxidant amount effective for inhibiting a decline in learning and/or memory, when added as an antioxidant composition to a food according to the present embodiment, will generally be found in a range as provided herein.
In a further embodiment, there is provided a method for inhibiting a decline in learning and/or memory in a dog comprising maintaining the dog on an antioxidant-fortified diet for a period of at least about 3 years, and wherein the diet comprises at least one antioxidant selected from the group consisting of vitamin E, vitamin C, α-lipoic acid and/or plant meal, the antioxidant being present in a total antioxidant amount effective to achieve such inhibition. The method of the invention will be found especially beneficial to a dog maintained on the fortified diet for an extended period of at least about 3 years, and preferably for the duration of the dog's life.
In a further aspect, the present invention provides kits suitable for administering one or more antioxidants to an animal. The kits comprise in separate containers in a single package or in separate containers in a virtual package, as appropriate for the kit component, at least one antioxidant and at least one of (1) one or more ingredients suitable for consumption by an animal, (2) instructions for how to combine the antioxidants and other kit components to produce a composition useful for inhibiting a decline in learning and/or memory in animals, and (3) instructions for how to use the antioxidants and other components of the present invention, particularly to inhibit a decline in learning and/or memory in animals. When the kit comprises a virtual package, the kit is limited to instructions in a virtual environment in combination with one or more physical kit components. The kit contains the antioxidants and other components in amounts sufficient to inhibit a decline in learning and/or memory in animals. Typically, the antioxidants and the other suitable kit components are admixed just prior to consumption by an animal. In one embodiment, the kit contains a packet containing one or more one or more antioxidants and a container of food for consumption by an animal. The kit may contain additional items such as a device for mixing the antioxidants and ingredients or a device for containing the admixture, e.g., a food bowl. In another embodiment, the antioxidants are mixed with additional nutritional supplements such as vitamins and minerals that promote good health in an animal.
In another aspect, the present invention provides a means for communicating information about or instructions for one or more of (1) using one or more antioxidants to inhibit a decline in learning and/or memory in animals, (2) admixing one or more antioxidants with the other components of the present invention, (3) administering one or more antioxidants to an animal, alone or in combination with the other elements of the present invention, and (4) using the kits of the present invention for inhibiting a decline in learning and/or memory in animals. The means comprises a document, digital storage media, optical storage media, audio presentation, or visual display containing the information or instructions. Preferably, the communication means is a displayed web site or a brochure, product label, package insert, advertisement, or visual display containing such information or instructions. Useful information includes one or more of (1) methods and techniques for combining and administering the antioxidants and/or other components and (2) contact information for animals or their caregivers to use if they have a question about the invention and its use. Useful instructions include amounts for mixing and administration amounts and frequency. The communication means is useful for instructing on the benefits of using the present invention and communicating the approved methods for administering the invention to an animal.
In a further aspect, the present invention provides for a use of a composition comprising a decline in learning and/or memory amount of at least one antioxidant to prepare a medicament. In another, the invention provides for the use of such composition to prepare a medicament for inhibiting a decline in learning and/or memory in animals. Generally, medicaments are prepared by admixing a compound or composition with excipients, buffers, binders, plasticizers, colorants, diluents, compressing agents, lubricants, flavorants, moistening agents, and other ingredients known to skilled artisans to be useful for producing medicaments and formulating medicaments that are suitable for administration to an animal. The medicament may further comprise one or more omega-3 fatty acids in amounts sufficient to inhibit a decline in learning and/or memory.
This invention is not limited to the particular methodology, protocols, and reagents described herein because they may vary. Further, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention. As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise, e.g., reference to “a method” or “a food” includes a plurality of such methods or foods. Similarly, the words “comprise”, “comprises”, and “comprising” are to be interpreted inclusively rather than exclusively.
Unless defined otherwise, all technical and scientific terms and any acronyms used herein have the same meanings as commonly understood by one of ordinary skill in the art in the field of the invention. Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred methods, devices, and materials are described herein.
All patents, patent applications, and publications mentioned herein are incorporated herein by reference to the extent allowed by law for the purpose of describing and disclosing the compounds, processes, techniques, procedures, technology, articles, and other compositions and methods disclosed therein that might be used with the present invention. However, nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.
EXAMPLE
The invention can be further illustrated by the following examples of preferred embodiments thereof, although it will be understood that these examples are included merely for purposes of illustration and are not intended to limit the scope of the invention unless otherwise specifically indicated.
Example 1
Seventeen young dogs of age 2-5 years were trained for six months on baseline learning and memory models in a standardized fashion. Group assignment of dogs to either a control food or an antioxidant-fortified food was based upon performance in these baseline models to assure that the dog assignments were balanced with respect to baseline level of cognitive function. The diet and methods of feeding that diet have been substantially described by Milgram et al. (2004) Experimental Gerontology 39(5), 753-765 and Milgram et al. (2005) Neurobiology of Aging 26(1), 77-90. Eight dogs were initially assigned to the control diet group and nine dogs to the antioxidant fortified diet. The antioxidant-fortified diet comprised basal dog food diet fortified with vitamin E, vitamin C, α-lipoic acid and fruit and vegetable meals. Table 1 shows the levels of vitamin E, vitamin C, α-lipoic acid and fruit and vegetable meals in the two diets.







TABLE 1







Comparison of Control Food to Antioxidant Fortified Food









INGREDIENT
CONTROL FOOD
FORTIFIED FOOD





Vitamin E
Appx 100 ppm
Appx 1000 ppm


l-Carnitine
None added
Appx 260 ppm


dl-α-lipoic acid
None added
Appx 120 ppm


Vitamin C
None added
Appx 80 ppm


Tomato Pomace*
None added
1%


Dried Spinach*
None added
1%


Dried Carrot*
None added
1%


Dried Citrus Pulp*
None added
1%


Dried Grape Pomace*
None added
1%





*Vegetables and Fruit Ingredients were added at 1% of the formula in exchange for corn.






The dogs were maintained on the assigned diet for 3 years. After 3 years on the control diet, three of six dogs passed a learning test involving simple shape discrimination while nine of nine dogs on the fortified diet passed the same learning test. The shape discrimination task involved presenting two wooden objects that are identical in thickness texture and color but different in shape. Subjects were given up to 40 training sessions to successfully complete the 2-stage learning criterion where a reward is given for the correct choice. The criterion is set as achieving a score of 8 of 10 correct for two consecutive days followed by a subsequent three days of an average of 70%. Once the initial discrimination is successfully achieved the reward for is reversed. Two dogs on the control diet and none on the fortified diet were dropped from the study for lack of motivation on the testing paradigm.
Table 2 illustrates the results of a memory test, delayed non-match to sample. None of the six dogs on the control diet passed the memory test while four of eight dogs on the antioxidant-fortified diet passed. This study showed that dogs on the antioxidant fortified diet retained memory for longer periods of time than the dogs on the control diet. The delayed non matching task consists of an initial presentation of the sample stimulus over the centre food well. Following a delay, a second presentation occurs with the sample stimulus presented along with a novel, second stimulus (delayed non match to sample or DNMS) At the four-year retest, only one of the animals in the control group was able to achieve criterion level of performance. Maximal memory is operationally defined as the longest delay interval that an animal could satisfactorily complete the two stage criterion as previously defined within 40 test sessions.







TABLE 2







Results of Memory Test









Mean maximal memory (seconds)










Diet group
Baseline
baseline + 1 year
baseline + 3 years













Control
40
10
0


Fortified
58
37
49









In the specification, there have been disclosed typical preferred embodiments of the invention and, although specific terms are employed, they are used in a generic and descriptive sense only and not for purposes of limitation, the scope of the invention being set forth in the following claims. Obviously many modifications and variations of the present invention are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims the invention may be practiced otherwise than as specifically described.

## Claims

1. A method for retaining memory as measured by a learning test involving the ability to discriminate simple shapes in an animal about 1 to about 6 years of age comprising maintaining the animal for at least three years on an antioxidant-fortified diet comprising: (a) antioxidants including at least 300 ppm of vitamin E, at least 10 ppm of vitamin C, and at least 25 ppm of α-lipoic acid based on a weight of the diet; and (b) at least one omega-3 fatty acid, wherein the animal is a young adult animal, and wherein the animal is a canine or a feline.

2. The method of claim 1 wherein the animal is the canine and the diet comprises a canned moist dog food or a dry dog food.

3. The method of claim 1 wherein the diet further comprises plant meal.

4. The method of claim 3 wherein the plant meal comprises fruit meal and/or vegetable meal.

5. The method of claim 1 wherein the diet is fortified with a nutritional supplement having the antioxidants as ingredients thereof.

6. The method of claim 1 wherein the antioxidants are distributed substantially homogeneously through a food component of the diet.

7. The method of claim 1 wherein the antioxidants are present in a food component of the diet in whole or in part on surfaces of food pieces.

8. The method of claim 1 further comprising adding to a food component of the diet an antioxidant composition that comprises a powder or liquid carrier having the antioxidant diluted or dispersed therein.