Note: Descriptions are shown in the official language in which they were submitted.
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COMPOSITIONS AND METHODS FOR REDUCING STOOL VOLUME AND STOOL ODOR
FIELD OF THE INVENTION
10001] The present invention relates to highly digestible pet food
compositions comprising
high quality protein and highly digestible carbohydrates and methods for
reducing pet stool
volume and pet stool odor by feeding an animal a composition of the present
invention,
BACKGROUND OF THE INVENTION
100021 Pets produce stool, and defecation may occur in any number of places,
e.g.,
outdoors, in litter boxes, or on newspaper. Cleaning up pet feces is often
unpleasant, and although
many products are marketed to make this task as pleasant as possible, the
products fail to address
the underlying problem of stool production, volume of stool produced, and
unpleasant odors
associated with stool. It is commonly known that food compositions can have a
great impact on
the amount of and characteristics of stool produced by an animal. Thus, it
would be desirable to
develop compositions and methods to reduce stool produced by an animal, either
by weight or
volume, and to reduce stool odor.
SUMMARY OF THE INVENTION
100031 The present invention relates to a highly digestible pet food
composition which
comprises high quality protein and highly digestible carbohydrates, the
ingestion of which causes
the formation of animal stools which are smaller in volume and have less
unpleasant odor
compared to stools produced by an animal fed compositions which contain lower
quantities of
high quality proteins and less digestible carbohydrates. Thus, in one aspect,
the invention relates
to a pet food composition comprising one or more high quality proteins, and
one or more highly
digestible carbohydrates. In certain embodiments, the compositions may include
from about 15%
to about 30% high quality protein and from about 25% to about 50% highly
digestible
carbohydrate. Said high quality protein may have a true protein digestibility
of greater than about
90%, 91%, 92%, 93%, 94%, or 95%; said highly digestible carbohydrate may have
a digestibility
of greater than about 90%, 92%, 94%, 95%, or 96%. The compositions of the
present invention
may have greater than about 88%, 90%, or 95% dry matter digestibility. In a
particular
embodiment, a composition of the present invention is a nutritionally complete
pet food, more
particularly, a dog food.
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[0003a] In a particular embodiment, the invention is directed to a highly
digestible
pet food composition comprising high quality protein and highly digestible
carbohydrates, wherein the pet food composition has a dry matter digestibility
greater
than about 88% and calculated as:
(mass of food consumed on a dry-matter basis - mass of stool produced) X 100%
mass of food consumed
[0004] In a particular embodiment, the invention is directed to a composition
comprising: 36% brewers rice, 16% egg, 16% corn starch, 14.5% corn gluten
meal,
2.5% beet pulp, 2.5% soybean oil, 2% flaxseed, carnitine, one or more amino
acids,
minerals and vitamins. Specifically,
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said composition displays ?88.0% dry matter, true protein, fat, carbohydrate,
and energy
digestibility.
[0005] In another aspect, the invention is directed to methods for reducing
the volume of stool
produced by an animal comprising feeding the animal any one of the highly
digestible
compositions of the present invention.
[0006] In a further aspect, the invention is directed to methods for reducing
the odor of stool
produced by an animal comprising feeding the animal any of one of the highly
digestible
compositions of the present invention.
[0007] In an additional aspect, the invention is directed to the use of high
quality proteins
and highly digestible carbohydrates in the manufacture of a food composition
to feed to an animal
in order to reduce the volume of stool produced by the animal.
[0008] In a further aspect, the invention is directed to the use of high
quality proteins and highly
digestible carbohydrates in the manufacture of a food composition to feed to
an animal in order to
reduce the odor of stool produced by the animal.
[0009] Other embodiments of the present invention will be apparent to those of
skill in the
art.
DETAILED DESCRIPTION OF THE INVENTION
[0010] Without being limited to any theories or particular modes of action of
the invention,
the present invention is based on the discovery that certain compositions,
when ingested by an
animal, can result in less stool production and the stool produced has a
reduced odor. Generally,
the compositions are highly digestible, comprising one or more high quality
proteins and highly
digestible carbohydrates. The increase in digestibility results in lower stool
volume produced. In
addition, it is believed that the reduction in stool odor is due to the
increase in digestibility and
associated reduction in the amount of sulfur containing compounds in the stool
produced by an
animal ingesting the compositions of the present invention.
[00111 As used herein and in the appended claims, the singular forms "a",
"an", and "the"
include plural reference unless the context clearly dictates otherwise.
[0012] As used herein, "an amount effective", "an effective amount", and like
terms refer to
that amount of a compound, material or composition as described herein that
may be effective to
achieve a particular biological result, e.g., to reduce stool volume and to
reduce stool odor. Such
result may be achieved, for example, by administration of the compositions of
the present
invention to an animal. An effective amount may be based on several factors,
including the
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particular animal, sex, age, weight, and/or the metabolizable energy of the
composition. As
contemplated herein, the daily amount of the food compositions to be fed to an
animal are familiar
to one of skill in the art.
[00131 As used herein, a "highly digestible diet" refers to a diet that has a
digestibility of
greater than or equal to 88.0%. The term "nutrient digestibility" includes the
digestibility of dry
matter, protein, fat, carbohydrate and energy.
100141 The present invention relates to any animal, preferably a mammal, more
preferably a
companion animal, which may benefit from feeding the formulations disclosed
herein. The term
"companion animal" refers to any animal that lives in close association with
humans and includes,
but is not limited to, pet canines and felines of any breed. For example, it
is contemplated herein
that this term may also encompass any animal whose diet may be controlled by
humans and which
may benefit from feeding the formulations disclosed herein. These animals may
include, e.g.,
domesticated farm animals (e.g. cattle, horses, swine, etc.) as well as
undomesticated animals held
in captivity, e.g. in zoological parks and the like.
100151 The present invention may be suitable for use with animals in various
stages in life,
including lactation, weaning, growth, adult, senior, and geriatric.
Preferably, the animal is an
adult, senior, or geriatric animal, preferably an adult animal.
[00161 Except to the extent stated otherwise, all percentages used in this
specification are
weight percentages on a dry matter basis. The phrase "dry matter basis" means
the component
concentration in the composition after any moisture in the composition is
removed.
100171 As referred to herein, a "high quality protein" is any protein or
mixture of proteins
that may be digested by a "true protein digestibility" (as opposed to
"apparent protein
digestibility") to greater than about 90%, preferably greater than about 95%,
more preferably
greater than about 98%.
100181 Methods for determining the digestibility of proteins are known by
those of skill in the
an. For example, protein content of compositions may be determined by any
number of methods
known by those of skill in the art, for example, as published by the
Association of Official Analytical
Chemists in Official Methods of Analysis ("OMA"), method 988.05. Thus, in
order to determine
"apparent protein digestibility", one of skill in the art would detemiine the
protein content of a
composition and the protein content of stool produced by feeding the animal
the composition. The
apparent protein digestibility is then calculated as follows:
(amount of protein consumed - protein content of stool) X 100%.
amount of protein consumed
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10019] It is known that stool contains many other protein sources, e.g.,
bacterial proteins,
enzymes produced by the digestion process, and bile. Thus, protein
digestibility may be skewed
by the presence of endogenous proteins in the stool. Thus, the apparent
protein digestibility is
often multiplied by a factor to take into account the presence of such
endogenous proteins to
determine a true protein digestibility percentage. Depending on the amount of
endogenous
proteins present, the factor may be increased or decreased. In the present
invention, the true
protein digestibility is determined by multiplying the protein digestibility
by a factor of 1.051.
That is, the difference between true protein digestibility and apparent
protein digestibility is 1.051
such that, e.g., an apparent protein digestibility of 88.5% would equate to
93.0% true protein
digestibility.
100201 Sources of high quality protein for use in the present invention
include plant sources,
animal sources, or both. Animal sources include, for example, meat, meat by-
products, seafood,
dairy products (including powdered milk), and egg. Vegetable sources may
include soy protein
isolate. High quality proteins can be intact, almost completely hydrolyzed,
partially hydrolyzed
proteins, or isolates. Preferably, the high quality protein is egg, e.g.,
chicken egg, substantially free of
shells. Other sources of high quality protein include egg white, casein,
hydrolyzed vegetable protein,
whey protein, ovalbumin, and lactalbumin. As contemplated herein, the
compositions of the present
invention may include any mixture of high quality proteins. Compositions of
the present invention
may comprise from about 15-30% of high quality protein.
10021] As referred to herein, "highly digestible carbohydrates" are those
carbohydrates that
may be digested by an animal, e.g., a canine, to greater than about 90%,
greater than about 93%,
preferably greater than about 95%, or greater than about 96%. Highly
digestible carbohydrates
may be supplied by any of a variety of sources known by those skilled in the
art, including corn,
wheat, rice (e.g. brewer's rice), starch (e.g., rice and corn starch), corn
gluten meal, and distiller's
dried grain. As contemplated herein, the compositions of the present invention
may include any
mixture of highly digestible carbohydrates. Compositions of the present
invention may comprise
from about 25-50% of highly digestible carbohydrates.
100221 Methods for detennining the digestibility of carbohydrates are known by
those of skill in
the art. Carbohydrate percentage may be calculated as nitrogen free extract
("NFE"), which may be
calculated as follows: NFE = 100% - moisture% - protein % - fat % - ash% -
crude fiber %. Thus, in
order to determine the carbohydrate digestibility, one of skill in the art
would determine the
carbohydrate content of a composition, and the carbohydrate content of stool
produced by feeding the
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animal the composition. The carbohydrate digestibility is then calculated by
the following equation:
(amount of carbohydrate consumed - carbohydrate content of stool) X 100%
amount of carbohydrate consumed
[0023] Dry matter digestibility (DMD) is the amount of matter that is digested
by an animal on a
dry matter basis. In general, digestibility is determined by measuring the
content ingested minus the
content in the feces divided by the content ingested. Methods for determining
DMD are known in the
art. For example, the mass of food consumed on a dry matter basis is
determined, and the mass of
stool (on a dry matter basis) resulting from consumption of the food is also
determined. DMD is then
calculated as follows:
(mass of food consumed - mass of stool produced) X 100%
mass of food consumed
100241 Fat and energy digestibility may also be determined as provided above,
e.g.,
(amount of fat consumed - fat content of stool) X 100%;
amount of fat consumed
(amount of energy consumed- energy content of stool) X 100%
amount of energy consumed
Determination of fat and energy content in food and stool may be performed
according to
conventional methods familiar to one of skill in the art.
[0025] In addition to causing a reduction in stool volume, the compositions of
the present
invention cause the stool produced by an animal ingesting said compositions to
be reduced in
odor. Methods for assaying the odor of stool are familiar to one of skill in
the art. For example,
qualitative assessment using human panelists such as described in the Examples
provided herein
may be used to rank stool odor. In addition, quantitative measurement of stool
odor compounds
using, e.g., gas chromatography and mass spectrometry may be used. Stool odor
compounds
quantitatively assayed in excreta include sulfur containing compounds,
phenols, heterocycles and
indoles, carboxylic acid compounds, and ketone compounds.
[0026] As contemplated herein, the compositions of the present invention are
meant to
encompass nutritionally complete pet food diets. A "nutritionally complete
diet" is a diet that
includes sufficient nutrients for maintenance of normal health of an intended
recipient animal on
the diet (e.g., domestic feline or domestic canine) and is familiar to one of
skill in the art. For
example, nutrients and ingredients such as those disclosed herein as well as
others suitable for
animal feed compositions, and recommended amounts thereof, may be found, for
example, in the
Official Publication of the Associate of American Feed Control Officials Inc.,
("AAFCO"),
Nutrient Requirements of Dogs and Ccits, 2006.
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100271 The nutritionally complete pet food compositions disclosed herein may
comprise fat.
Sources of fat for the compositions of the present invention can be supplied
by any of a variety of
sources known by those skilled in the art, including meat, meat by-products,
fish oil, and plants.
Plant fat sources include wheat, flaxseed, rye, barley, rice, sorghum, corn,
oats, millet, wheat
germ, corn germ, soybeans, peanuts, and cottonseed, as well as oils derived
from these and other
plant fat sources. As contemplated herein, the compositions of the present
invention may
comprise from about 11 % to 16% fat, preferably about 14% fat.
100281 Vitamins and minerals may also be included in the compositions
described in the
instant invention in amounts known to those of skill in the art to avoid
deficiency and maintain
health. For example, the National Research Council (NRC) provides recommended
amounts of
such ingredients for farm animals, and AAFCO (e.g., Official Publication of
the Associate of
American Feed Control Officials Inc., ("AAFCO"), Nutrient Requirements ofDogs
and Cats,
2006) provides recommended amounts of such ingredients for canines and
felines. Vitamins may
include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C,
vitamin D, vitamin E,
vitamin H (biotin), vitamin K, folic acid, inositol, niacin, and pantothenic
acid. Examples of
minerals include e.g., calcium, phosphorus, dicalcium phosphate, potassium
citrate, potassium
chloride, calcium carbonate, sodium chloride, or salts of sodium, potassium,
magnesium, manganese,
calcium, chloride, copper, zinc, choline, phosphorus, iodine or selenium.
100291 Fiber, including but not limited to dietary fiber, may be included in
the compositions
of the present invention. Dietary fiber refers to components of a plant that
are resistant to
digestion by an animal's digestive enzymes. Dietary fiber components of foods
may be determined
by any number of methods known by those of skill in the art, such as OMA
method
991.43/32.1.17 (1994). Total dietary fiber includes soluble and insoluble
fibers. Soluble fiber is
resistant to digestion and absorption in the small intestine, but undergoes
complete or partial
fermentation in the large intestine. Sources of soluble fiber for use in the
compositions disclosed
herein include, e.g., beet pulp, guar gum, chicory root, psyllium, pectin,
blueberry, cranberry,
squash, apples, oats, beans, citrus, barley, or peas. Insoluble fibers are
fibers that do not dissolve
in water and tend to increase the rate at which food passes through the
digestive tract. Examples
include, but are not limited to, cellulose, whole wheat products, wheat oat,
corn bran, flax seed,
grapes, celery, green beans, cauliflower, potato skins, fruit skins, vegetable
skins, peanut hulls, and
soy fiber. As contemplated herein, the compositions of the present invention
may comprise from
about 3% to 6% total dietary fiber.
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[0030] The compositions of the present invention may additionally comprise
amino acids in
amounts known to those of skill in the art to avoid deficiency and maintain
health. Preferably, the
compositions of the present invention comprise "essential amino acids."
Essential amino acids are
amino acids that cannot be synthesized de novo, or in sufficient quantities by
an organism and
thus must be supplied in the diet. Essential amino acids vary from species to
species, depending
upon the organism's metabolism. For example, it is generally understood that
the essential amino
acids for dogs and cats (and humans) are phenylalanine, leucine, methionine,
lysine, isoleueine,
valine, threonine, tryptophan, histidine and arginine. In addition, taurine,
while technically not an
amino acid but a derivative of cysteine, is an essential nutrient for cats and
dogs.
[0031] The compositions of the present invention may additionally comprise
nutritional
supplements, e.g., carnitine and omega-3 or omega-6 fatty acids, in amounts
known to those of
skill in the art to avoid deficiency and maintain. health. Carnitine, or L-
carnitine, is a vitamin-like
compound synthesized in the body from lysine and methionine. Carnitine may be
naturally present
in the ingredients of the present invention (e.g., flaxseed), or camitine may
be added to the
compositions.
[0032] The compositions of the present invention may additionally comprise
additives,
stabilizers, fillers, thickeners, flavorants, palatability enhancers and
colorants in amounts and
combinations familiar to one of skill in the art. Stabilizing substances may
increase the shelf life of
the composition, and are known by those skilled in the art.
[0033] As contemplated herein, the compositions of the present invention may
comprise a
metabolizable energy content of from about 3500 to about 4500 kcal/kg. As
referred to herein,
"metabolizable energy" is the energy available to an animal upon consumption
of the diet (or
composition) after subtracting the energy excreted in feces, urine, and
combustible gases.
Metabolizable energy values may be determined following the protocols
established by methods
known by those of skill in the art, e.g., according to official guidelines
produced by the
Association of American Feed Control Officials (AAFCO).
[0034] By way of example, but without limitation, it is contemplated herein
that a highly
digestible composition of the present invention which results in reduced stool
volume and stool
malodor may comprise: 36% brewers rice, 16% egg, 16% corn starch, 14.5% corn
gluten meal,
2.5% beet pulp, 2.5% soybean oil, 2% flaxseed, carnitine, one or more amino
acids, minerals and
vitamins and one or more palatability enhancers. Specifically, said
composition displays ?88.0%
dry matter, true protein, fat, carbohydrate, and energy digestibility.
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[0035] While compositions of any consistency or moisture content are
contemplated,
preferably the compositions of the present invention may be, for example, a
wet, semi-moist, or
dry animal food composition. "Wet" food refers to food that has a moisture
content of from about
70 to 90%. "Semi-moist" food refers to food that has a moisture content of
from about 15% to
40%, "Dry" food refers to compositions from about 5% to 15% moisture content
and is often
manufactured in the form of small bits or kibbles. Also contemplated herein
are compositions that
may comprise components of various consistency as well as components that may
include more
than one consistency, for example, soft, chewy meat-like particles as well as
kibble having an
outer cereal component and an inner cream component as described in, e.g., US
Patent 6,517,877.
The kibble may then be dried and optionally coated with one or more topical
coatings known by
those skilled in the art, for example, flavors, fats, oils, powders, and the
like. The compositions of
the present invention can be prepared using conventional manufacturing
methods.
[0036] It is contemplated that the invention described herein is not limited
to the particular
methodology, protocols, and reagents described as these may vary. It is also
to be understood that
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 in any way. Further,
the detailed
description of embodiments is intended only to acquaint others skilled in the
art with the
invention, its principles, and its practical application so that others
skilled in the art may adapt and
apply the invention in its numerous forms, as they may be best suited to the
requirements of a
particular use. This invention, therefore, is not limited to the embodiments
described herein, and
may be variously modified.
[0037] Unless defined otherwise, all technical and scientific terms used
herein have the same
meanings as commonly understood by one of ordinary skill in the art to which
this invention
belongs.
EXAMPLES
Example 1
[0038] The foods of Table 1 are formulated and formed as kibbles in accordance
with the
Association of American Feed Control Officials 2005 Nutrient Guide for Dogs,
balanced to meet
adult maintenance requirements and extruded as a dry kibble according to
conventional methods.
Both compositions are formulated to meet or exceed AAFCO nutrient
recommendations.
Metabolizable energy is calculated according to conventional methods, e.g.,
using the Atwater
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equation, and percentages are expressed on a dry matter basis. The five most
abundant ingredients
(in decreasing order) are also provided:
Table 1: Nutrient Analysis and Top Five Ingredients of Control A and Test
Composition B
Analyzed nutrient (dry matter basis) Control A Test Composition B
Crude protein % 25 24
Crude fat % 15.5 12.2
Calcium % 0.74 0.77
Phosphorus % 0.67 0.5
Crude fiber % 2.2 1.1
Metabolizable Energy (kcal/kg) 4035 3900
Lysine, % 1.14 1.37
Lysine:calorie 2.8 3.5
Ingredient
1 Corn Rice
2 Milo Egg
3 Wheat Corn Starch
4 Ground chicken Corn gluten meal
Poultry Meal Beet pulp
Example 2
[0039] A digestibility study is performed to determine the digestibility of
the control and
test foods of Example 1. Two sets of five dogs over the course of 14 days are
fed the
compositions of Example 1. One set of five dogs is fed Control A, and the
second set of five dogs
is fed Test Composition B for seven days. Following seven days, the food for
the animals is
exchanged. Days 1 and 2 are allowed for adaptation to the food, and on days 3
to 7 total stool
amounts are collected and analyzed. On day eight, each group is fed the second
food, the first two
days allowing for adaptation to the new food, and stool samples are collected
daily and analyzed
for the remainder of the study. Feces are collected throughout the day and
placed in sealable
plastic bags (24 hr or before cage washing the following morning). Bags are
labeled with animal
ID #, date and test #. Any fecal sample needing storage is refrigerated at 5
C. Fecal volume is
measured using a displacement method. The stool sample is weighed and then
added at the top of
a stool densitometer. The displaced water is collected and the volume measured
in a graduated
cylinder. The density measurement (g/cc) is the following calculation: the
weight of the sample
(grams) divided by the volume (mL) of the displaced water. Data are analyzed
using the General
Linear Models procedure of SAS to determine treatment means. All digestibility
assays are
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performed according to conventional methods. Digestibility values, and stool
characteristics are
provided in Table 2.
Table 2: Results of Digestibility Study Comparing Control A with Test
Composition B
Test Result Control A Test Composition B
Dry matter digestibility % 86.2 89.5
True Protein digestibility % 89.2 93.8
Fat digestibility % 94.5 94.2
Carbohydrate digestibility % 95.5 96.0
Energy digestibility % 88.6 92.4
Stool quantity (g/day) 119.2 87.8
Average stool weight (g) 95.3 64.0
Average stool volume (ml) 89.8 63.1
Average Stool Density (g/ml) 1.06 1.01
Average daily food intake (g) 300 298
[0040] The results indicate that animals fed Test Composition B produce less
stool (in
terms of stool weight and volume) compared to dogs fed Control A.
Example 3
[0041] Test Composition C is formulated in accordance with the Association of
American
Feed Control Officials 2005 Nutrient Guide for Dogs, balanced to meet adult
maintenance
requirements, and extruded as a dry kibble according to conventional methods.
Metabolizable
energy content is calculated with the Atwater equation. The four most abundant
ingredients are
rice, egg, corn starch, and corn gluten meal. The contents of Test Composition
C are analyzed by
methods known in the art, and are presented below on a dry matter basis.
Table 3: Analyzed Nutrients of Test Composition C
Nutrient Test Composition C
Crude Protein, % 25.1
Crude Fat, % 13.8
Calcium, % 0.64
Ash % 5.1
Phosphorous, % 0.62
Crude Fiber, % 0.90
NFE% 55.1
Total Dietary Fiber, % 4.65
Metabolizable Energy, kcal/kg 3978
Example 4
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100421 A digestibility study is performed to compare Test Composition C of
Example 3
with a commercially available dog food (Eukanuba Adult Maintenance Small
Bites ("EU-
SB")). Metabolizable energy content is calculated with the Atwater equation.
The four most
abundant ingredients of EU-SB are chicken, chicken-by-product meal, corn meal,
and ground
whole grain sorghum. The composition of EU-SB is analyzed by methods known in
the art, and is
provided in Table 4 along with the nutrient analysis of Test Composition C.
The results of the
digestibility study are presented in Table 5.
100431 The digestibility study is performed as described in Example 2. In this
case, two
sets of five dogs over the course of 14 days are fed either Test Composition C
or EU-SB. One set
of five dogs are fed Composition C, and the second set of five dogs are fed EU-
SB for seven days.
Following seven days, the foods for the animals are exchanged. Days l and 2
are allowed for
adaptation to the food, and on days 3 to 7 total stool amounts are collected.
On day eight, each
group is fed the second food, the first two days allowing for adaptation to
the new food. Stool
samples are collected daily and analyzed for the remainder of the study as
described in Example 2.
With regard to each 7 day test period, the first 3 day stool collection is
used for measurement of
stool volume, weight and density. These three stool measurements are assessed
daily and a three
day average is computed. Additionally in this study, the last two days of the
stool collection are
used for measurement of fecal odor. A two day composite is used for the fecal
odor measurement
(see Example 5 below). A study schedule is provided below:
Study Day Procedure Measurement
Days 0-7 5 animals receive Control Food, 5 Food Intake & Fecal Score- daily
animals receive Test Food Body Weight -weekly
Day 3-5 Fecal collection Fecal volume, weight & density
Day 6-7 Fecal collection Fecal odor
Days 8-14 Animals switch diet; 5 on control, 5 Food Intake & Fecal Score-daily
on test
Day 10-12 Fecal collection Fecal volume, weight & density
Day 13-14 Fecal collection Fecal odor
Table 4 - Analyzed Nutrients of Food Compositions
Nutrient (on dry matter basis) Test Composition C EU-SB
Crude Protein % 25.1 26.5
Crude Fat % 13.8 18.2
Ash % 5.1 7.7
Calcium % 0.64 1.35
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Phosphorous % 0.62 1.07
Crude Fiber % 0.90 2.35
NFE % 55.1 45.2
Total Dietary Fiber % 4.65 9.74
Metabolizable Energy (kcal/kg) 3978 4058
Table 5: Results of Digestibility Study Comparing Test Composition C and EU-SB
Test Result Test Composition C EU-SB
Dry Matter Digestibility % 91.0 84.9 a
True Protein Digestibility % 98.2 89.9 a
Fat Digestibility % 94.3 95.5 a
Carbohydrate Digestibility % 96.7 95.9
Energy Digestibility % 92.7 89.1 a
Stool volume (ml) 54.7 90.1
Stool weight (g) 57.4 95.6
Stool density (g/cc) 0.99 1.05
a p < 0.05
b p < 0.10
[0044] The results indicate that when Test Composition C is fed to dogs, the
dogs produce
less stool weight and volume than dogs fed commercial food EU-SB.
Example 5
[0045] Fecal odor analyses are performed with the stools collected from the
digestibility
study described in Example 4. All fecal samples are kept refrigerated at 5 C
prior to analysis.
The samples are removed from refrigeration and composited by animal as needed
into
polyethylene bags. The samples are then kneaded in the bag to break up and mix
the fecal
material. Fecal material is then loaded into a lOmL polyethylene syringe and
5g (+/- 0.01 g) is
expressed into a 20mL headspace vial and crimp capped for analysis. Expression
from the syringe
serves to increase the surface area of the sample. This is critical for
adequate equilibration of the
volatile components in the stool samples. Sample preparation in this manner
also serves to
"normalize" the form of the samples in the headspace vial.
[0046] The samples are then loaded onto a Leap Technologies Combi-PAL sampler.
The
sampler took each sample, in turn, and incubated them at 30 C and then sampled
using a Solid
Phase Microextraction (SPME) fiber (Supelco, 1 cm x 75 gin Carboxen/PDMS). The
fiber is
exposed to the headspace in the vial for 30 min and then injected onto an
Agilent 6890 Gas
Chromatograph/5973N Mass Selective Detector (GCMS). The separation is done on
an Agilent
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DB-WAXETR column (60m x 0.25mm x 0.25 m). When the analysis is complete, the
data is
processed to measure the peak area for an ion (the Qlon) unique to each
compound.
[0047] A summary report of all the peak areas is exported into a database
where each
compound is categorized by its primary functional group. The peak areas of all
compounds in
each category are totaled. These totals are then analyzed using the SAS pair-
wise comparisons
Proc mixed procedure.
[0048] Stool aroma evaluations are made on study days 6, 7, 13, and 14 using
the
following procedures: fresh stool samples are collected and placed in plastic
bags that have been
randomly coded with 3-digit numbers. Eleven animal colony staff members are
recruited to
evaluate the samples. Samples are evaluated within 1 hour of collection.
Samples are given, one
at a time, to each participant in random order. Participants evaluate the
samples by holding the
open bag approximately 1 foot from their nose and smell the aroma using
several "bunny sniffs".
Participants assign an aroma strength rating to each sample using a 5-point
scale where l=no
aroma at all and 5=extremely strong aroma. Aroma strength ratings are recorded
on forms
provided. At the end of the study, means for each sample are calculated and
compared using the
general linear model (GLM) procedure of SAS. Results of the fecal analyses are
provided below
in Tables 6-14.
Table 6: Fecal Stool Scores
EU-SB
Intake Data:
Mean StDev Min Max Count
Food Intake 223.5 33.9 1.29 303 95
Stool 4.9 .3 4 5 97
Stool Frequency Data:
Stool Rating 1 2 3 4 5
Count 0 0 0 10 87
Percent 0.0 0.0 0.0 10.3 89.7
Test Composition C
Intake Data:
Mean StDev Min Max Count
Food Intake 213.3 30.4 157 277 95
Stool 4.9 .4 2 5 93
Stool Frequency Data:
Stool Rating 1 2 3 4 5
Count 0 1 1 5 86
Percent 0.0 1.1 1.1 5.4 92.5
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EU-SB
Intake Data:
Mean StDev Min Max Count
Food Intake 181.3 41.1 124 275 70
Stool 4.3 .7 2 5 101
Stool Frequency Data:
Stool Rating 1 2 3 4 5
Count 0 2 10 41 48
Percent 0.0 2.0 9.9 40.6 47.5
Test Composition C
Intake Data:
Mean StDev Min Max Count
Food Intake 181.3 41.6 123 277 70
Stool 4.4 .7 2 5 79
Stool Frequency Data:
Stool Rating 1 2 3 4 5
Count 0 2 6 29 42
Percent 0.0 2.5 7.6 36.7 53.2
Table 7: Average Fecal Volume, Weight and Density
Fecal Weight (grams) Fecal Volume (mL) Fecal Density (g/cc)
EU-SB 95.60 90.10 1.05
Test Comp. C 57.40 54.70 0.99
P-value < 0.01 < 0.01 0.03
% Difference -39.96% -39.29% -5.71%
Table 8 - Fecal Odor: Comparison of Peak Area Totals for Classes of Headspace
Volatiles
from Canine Stools for Dogs on Test Composition C and EU-SB - Based on Overall
Feeding
- (Statistics using Pairwise Comparisons by Proc Mixed) - Test I
Test Composition C EU-SB Probability of > t
Acids 20210973 38915911 0.177
Alcohols 14099355 14946040 0.849
Ketones 14278852 11038781 0.167
Aldehydes 793065 723638 0.654
Sulfur-Containing * 2578798 7415707 0.084
Heterocyclics* 6164309 3963024 0.070
Furans# 1109634 587942 0.017
Esters 116421 11 16152892 0.244
Indole 4542689 3017321 0.141
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Phenol* 3748062 2047024 0.094
# Peak areas are significantly different at the 95% confidence level
* Peak areas are significantly different at the 90% confidence level
Peak areas are adjusted for sample weights
Table 9 - Fecal Odor: Comparison of Peak Area Totals for Classes of Headspace
Volatiles
from Canine Stools for Dogs on Test Composition C and EU-SB- Based on Overall
Feeding
- (Statistics using Pairwise Comparisons by Proc Mixed) - Test 2
Test Composition C EU-SB Probability of > t
Acids* 27576089 54175201 0.0211
Alcohols# 9834550 7401144 0.0987
Aldehydes* 686314 171381 0.0020
Esters 10082059 12901126 0.2351
Furans 783506 624359 0.3247
Heterocyclics* 3490767 2120497 0.0153
Hydrocarbons* 165010 328482 0.0046
lndole* 3147117 1829258 0.0200
Ketones 11546838 15886302 0.5580
Phenol* 2484498 1253450 0.0334
Sugar Related* 769426 4875788 0.0079
Sulfur Compounds 1073366 2497176 0.3220
* Peak areas are significantly different at the 95% confidence level
# Peak areas are significantly different at the 90% confidence level
Peak areas are adjusted for sample weights
Table 10: Comparison of Peak Area Totals between Studies (Tests 1 and 2);
Directional
Changes in Peak Area for Test Composition C vs EU-SB
Test 1 Test 2
Acids I I
Alcohols T
Aldehydes T
Esters I I
Furans T T
Heterocyclics T T
Indole T
Ketones 1
Phenol T
Sulfur Compounds 4 1
Table 11 - Fecal Odor: Comparison of Peak Area Totals for Classes of Headspace
Volatiles
from Canine Stools for Dogs on Test Composition C and EU-SB - Based on Overall
Feeding
- (Statistics using Pairwise Comparisons by Proc Mixed - Test 1 and Tests 2
Combined
I Test Composition C EU-SB Probability of > Iti
Acids* 23893531 46545556 0.0102
Alcohols 11966953 11173592 0.6598
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Aldehvdes# 739690 447510 0.0745
Esters# 10862085 14527009 0.0905
Furans* 946570 606151 0.0264
Heterocyclics# 4827538 3041761 0.0525
Indole* 3844903 2423290 0.0455
Ketones 12912845 13462542 0.8854
Phenol* 3116280 1650237 0.0456
Sulfur Compounds* 1826082 4956442 0.0490
* Peak areas are significantly different at the 95% confidence level
# Peak areas are significantly different at the 90% confidence level
Peak areas are adjusted for sample weights
Table 12: Fecal Sensory Human Assessment: Aroma Strength Scores for Dogs Fed
Dry -
Test Composition D and EU-SB- Total Test (Test 2)
1 Test Composition C EU-SB
Aroma Strength 2.92 a 3.19
Distribution of Odor Scores
1=No Aroma at All 7% 1%
2=Slight Aroma 31% 23%
3=Moderate Aroma 32% 39%
4=Strong Aroma 18% 24%
5=Extremely Strong Aroma 10% 11%
a Means followed by the same letter followed by the same letter are not
significant at the
95% confidence level
Table 13: Fecal Sensory Human Assessment: Aroma Strength Scores for Dogs Fed
Test -
Composition C and EU-SB- Grouped by Feeding Group (Test 2)
Group 1 Group 2
Feeding EU-SB First - Test Composition C First- EU-SB
Sequence Test Composition C Last Last
Food Test Comp. EU-SB Test Comp. EU-SB
Aroma 2.74 a 3.28 b 3.11 a 3.10 a
Strength I
Distribution of Odor Scores
None 9% 1% 5% 1%
Slight 37% 21% 25% 24%
Moderate 32% 37% 32% 42%
Strong 11% 24% 25% 23%
Extreme 7% 14% 11% 7%
a Means followed by the same letter within a group that are followed by the
same
letter are not significant at the 95% confidence level
Table 14 - Fecal Sensory Human Assessment: Aroma Strength Scores for Dogs Fed
Test
Composition C and EU-SB - Grouped by Feeding Sequence (Test 2)
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Group I Group 2
Feeding EU-SB First - Test Composition C - First EU-SB
-Sequence Test Composition C Last Last
Date 3/1 2/22 3/2 2/23 2/22 3/1 2/23 3/2
Food Test EU-SB Test EU-SB Test EU-SB Test EU-SB
Aroma 2.73 a 3.33b 2.76 a 3.24b 2.98 a 3.02a 3.24 a 3.20 a
Strength
Distribution of Odor Scores
None 12% 1% 6% 1.% 9% 3% 1% -
Slight 34% 14% 40% 29% 25% 27% 25% 21%
Moderate 32% 45% 32% 29% 30% 40% 30% 45%
Strong 7% 25% 16% 23% 27% 21% 27% 25%
Extreme 12% 12% 6% 16% 7% 7% 7% 7%
n a Means followed by the same letter within a comparison that are followed by
the same letter are
not significant at the 95% confidence level
[0049] Data presented in the tables above indicate that fecal stool scores are
not different
between Test Composition C and EU-SB (e.g., 4.9 for both diets in Test 1; 4.3
vs 4.4 for EU-SB
and Test Composition C, respectively in Test 2).
[0050] Fecal stool weights and volumes are statistically different between
Test
Composition C and EU-SB product. In both studies (Test I and Test 2), dogs fed
the test diet
have significantly lower stool weight (57.4 vs 95.6 g, respectively) and lower
stool volume (54.7
vs 90.1 mL, respectively) compared to dogs fed the EU-SB product. Fecal
density is not different
(0.99 vs 1.05 g/cc). Numbers shown are an average from the two studies.
100511 Fecal odor is assessed using headspace analysis of the stool aroma.
Results from
the combined studies show that dogs fed the test diet produce 63% lower levels
of sulfur-
containing compounds compared to dogs fed the EU-SB product (P < 0.05). This
is an important
finding because sulfur compounds have a very low odor threshold, thus a
reduction in these
compounds should result in an improvement in stool odor. Dogs fed the test
diet also have
significantly lower levels of acids (e.g., 49% reduction; (P < 0.05)). Dogs
fed the EU-SB diet
(numbers shown are from the combined study) have significantly lower levels of
phenols (P <
0.05), indoles (P < 0.05), and furans (P < 0.05).
[0052] A sensory panel composed of 11 participants is used to rank stool odor
strength in
stool samples collected from the study 2 stool samples. These analyses show
that dogs fed the test
diet had a lower fecal odor score compared to dogs fed the EU-SB product (2.92
vs 3.19; P <
0.05). Thus, viewing the results as a whole (e.g., the headspace analyses
along with the results
from the sensory analyses) suggests that reductions in sulfur and possibly
acid compounds may be
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more important than reductions in some of the other compounds (e.g., furans,
phenols, and
indoles).
10053 The Test Composition C dog diet is shown to be very highly digestible.
Digestibility of dry matter, true protein and energy is 91.0, 98.2 and 92.7%,
respectively. Not
only does the increased digestibility reduce stool weight and volume, but
fecal odor (as assessed
by both analytical and sensory tests) is also reduced.
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