U.S. patent application number 15/870199 was filed with the patent office on 2018-05-17 for composition and method for promoting reduction of heat stress in animals.
This patent application is currently assigned to OmniGen Research, L.L.C.. The applicant listed for this patent is OmniGen Research, L.L.C.. Invention is credited to David Calabotta, James D. Chapman, Neil E. Forsberg, Steven B. Puntenney.
Application Number | 20180133242 15/870199 |
Document ID | / |
Family ID | 52598820 |
Filed Date | 2018-05-17 |
United States Patent
Application |
20180133242 |
Kind Code |
A1 |
Chapman; James D. ; et
al. |
May 17, 2018 |
COMPOSITION AND METHOD FOR PROMOTING REDUCTION OF HEAT STRESS IN
ANIMALS
Abstract
Disclosed herein is a method for promoting reduction of heat
stress in animals, as well as a composition for use in the
disclosed method. The composition comprises a glucan, silica,
mineral clay, mannan, and optionally an endoglucanohydrolase, and
may be administered to an animal that is susceptible to or suffers
from heat stress.
Inventors: |
Chapman; James D.; (Macon,
GA) ; Calabotta; David; (Quincy, IL) ;
Forsberg; Neil E.; (Corvallis, OR) ; Puntenney;
Steven B.; (Ione, OR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
OmniGen Research, L.L.C. |
Corvallis |
OR |
US |
|
|
Assignee: |
OmniGen Research, L.L.C.
Corvallis
OR
|
Family ID: |
52598820 |
Appl. No.: |
15/870199 |
Filed: |
January 12, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15234971 |
Aug 11, 2016 |
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15870199 |
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PCT/US2015/015692 |
Feb 12, 2015 |
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15234971 |
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62000986 |
May 20, 2014 |
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61939206 |
Feb 12, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A23K 50/40 20160501;
A23K 50/20 20160501; A23K 20/20 20160501; A61K 31/736 20130101;
A61K 33/06 20130101; A61K 45/06 20130101; A61K 38/47 20130101; A61K
31/716 20130101; A61K 35/00 20130101; A23K 50/70 20160501; A61K
35/02 20130101; A23K 20/163 20160501; A23K 20/28 20160501; A61K
33/00 20130101; A23K 50/10 20160501; A23K 50/30 20160501; A61K
31/715 20130101; A61K 31/716 20130101; A61K 2300/00 20130101; A61K
33/00 20130101; A61K 2300/00 20130101; A61K 31/715 20130101; A61K
2300/00 20130101; A61K 35/02 20130101; A61K 2300/00 20130101; A61K
38/47 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/716 20060101
A61K031/716; A23K 50/70 20060101 A23K050/70; A23K 50/30 20060101
A23K050/30; A61K 31/715 20060101 A61K031/715; A61K 33/00 20060101
A61K033/00; A61K 31/736 20060101 A61K031/736; A61K 33/06 20060101
A61K033/06; A61K 35/00 20060101 A61K035/00; A23K 20/163 20060101
A23K020/163; A23K 20/20 20060101 A23K020/20; A23K 20/28 20060101
A23K020/28; A61K 38/47 20060101 A61K038/47; A23K 50/40 20060101
A23K050/40; A23K 50/10 20060101 A23K050/10; A23K 50/20 20060101
A23K050/20; A61K 35/02 20060101 A61K035/02; A61K 45/06 20060101
A61K045/06 |
Claims
1. A method, comprising: administering to an animal that has or is
at risk of developing heat-induced stress a composition comprising
glucan, silica, mineral clay, and mannan, wherein the animal is
other than a cow or a fish.
2. The method of claim 1, wherein the animal is an avian, a
non-ruminant mammal, or a ruminant mammal selected from the group
consisting of sheep, goats, deer, bison, buffalo, or llama.
3. The method of claim 2, wherein the animal is an avian, and the
avian is a chicken, turkey, goose, duck, cornish game hen, quail,
pheasant, guinea-fowl, ostrich, emu, swan, or pigeon.
4. The method of claim 2, wherein animal is a non-ruminant animal,
and the non-ruminant mammal is a dog, cat, rabbit, horse, donkey,
or pig.
5. The method of claim 2, wherein the animal is a ruminant mammal
selected from the group consisting of sheep, goats, deer, bison,
buffalo, or llama.
6. The method of claim 1 wherein the composition comprises between
15% and 40% silica, between 50% and 81% mineral clay, between 1.0%
and 5.0% .beta.-glucans, and between 1% and 8.0% mannan.
7. The method of claim 1, the composition is administered to the
animal prior to the animal experiencing heat stress, while the
animal is experiencing heat stress, and after the animal
experiences heat stress.
8. The method of claim 1, wherein the composition is administered
for an effective period of time to promote reduction of heat
induced stress.
9. The method of claim 8, further comprising administering the
composition to the animal at set intervals for the effective period
of time.
10. The method of claim 8, wherein the effective period of time is
at least 1 day.
11. The method of claim 1, further comprising administering the
composition to the animal when a temperature humidity index is, or
is expected to be, greater than 68.
12. The method of claim 11, further comprising: increasing an
amount of the composition administered to the animal if the
temperature humidity index is expected to increase; or decreasing
an amount of the composition administered to the animal if the
temperature humidity index is expected to decrease.
13. The method of claim 1, further comprising administering to the
animal an amount of the composition effective to alter a heat
stress indicator compared to an animal that has not been
administered the composition.
14. The method of claim 13, wherein the heat stress indicator is
feed intake, water consumption, respiration rate, rectal
temperature, milk yield, milk fat, milk protein, an immune
biomarker, or any combination thereof.
15. The method of claim 13, further comprising: measuring the heat
stress indicator; and selecting an amount of the composition
administered to the animal based at least in part on the
measurement of the heat stress indicator.
16. The method of claim 13, further comprising: administering a
first amount of the composition daily to the animal for a first
period of time; measuring the heat stress indicator; and
administering an adjusted amount of the composition daily to the
animal for a subsequent period of time, wherein the adjusted amount
is based at least in part on the measurement of the heat stress
indicator.
17. The method of claim 1, wherein administering comprises mixing
the composition with the animal's feed in an amount ranging from
0.1 to 100 kg per ton of feed, and providing the composition mixed
with the feed to the animal.
18. The method of claim 1, further comprising administering to the
animal a therapeutic process for alleviating heat stress, a
therapeutic agent for treating heat stress, or a combination
thereof.
19. The method of claim 18, wherein the therapeutic process or
therapeutic agent is provision of shade to the animal, use of a
water sprinkler to externally administer water to the animal, use
of a fan to provide air movement, addition of bypass fats to feed
for the animal, meloxicam, a corticosteroid, a composition
comprising one or more electrolytes, an alkalinizing agent, a
direct-fed microbial, or a combination thereof.
20. The method of claim 1, further comprising: evaluating at least
one heat stress indicator of individual animals in a group of
animals; determining whether one or more of the individual animals
are experiencing heat stress; and administering the composition to
the one or more individual animals that are experiencing heat
stress.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This is a divisional of U.S. application Ser. No.
15/234,971, filed Aug. 11, 2016, which is a continuation of
International Application No. PCT/US2015/015692, filed Feb. 12,
2015, which claims the benefit of U.S. Provisional Application No.
62/000,986, filed May 20, 2014, and the benefit of U.S. Provisional
Application No. 61/939,206, filed Feb. 12, 2014, each of which is
incorporated by reference herein in its entirety.
FIELD
[0002] This disclosure relates to methods and compositions for
promoting reduction of heat stress in animals.
BACKGROUND
[0003] Heat stress causes significant economic losses in animal
industries, such as dairy industries. Production, reproduction and
animal health are impaired by hyperthermia. Physiological and
production responses are well documented, but not fully understood.
During heat stress, respiration rates and body temperatures
increase, and feed intake, milk yield, and reproduction
decrease.
[0004] Milk synthesis decreases from reduced feed intake. There are
additional losses in milk yield that are associated metabolic
changes (Rhoads et al., 2009). Countering the production effects of
heat stress in lactating dairy cows involves reduced maintain feed
intake and controlling metabolic flux.
[0005] Immune function and health are also reduced with heat
stress. The severity and occasion of disease are increased when
immune and inflammatory responses are impaired. The rumen
environment can be altered during thermal stress. Increased
respiration rates can cause respiratory alkalosis, rumen acidosis
and eventually metabolic acidosis. Oxidative stress can be
increased with heat stress and can impair the heat shock response
and increase cell damage and death.
SUMMARY
[0006] The object of the present disclosure is to provide a novel
and previously unknown method for promoting reduction of heat
stress in animals. Also disclosed herein is a composition suitable
for administering to animals in order to substantially prevent
and/or promote reduction of heat stress in animals. The disclosed
composition and method for preventing and/or promoting reduction of
heat stress may be applied to any animal species susceptible to
heat stress, such as mammals, avians, or aquatic animal species. In
some embodiments, the animals treated herein can be an animal
raised for human consumption, such as livestock (e.g., feed or
dairy cattle) or pigs, or avians, such as domestic fowl (e.g.,
chicken, turkey, goose, duck, cornish game hen, quail, pheasant,
guinea-fowl, ostrich, emu, swan, or pigeon), or fish (e.g., salmon,
trout and the like). In other embodiments, the animal can be a
domestic animal, such as a dog, cat, fish, or rabbit. In some other
embodiments, the animal can be a ruminant species, such as a sheep,
goat, cow, deer, bison, buffalo, or llama. In yet other
embodiments, the animal can be an ungulate, such as a horse,
donkey, or pig.
[0007] Disclosed herein are embodiments of a method for promoting
heat stress reduction in an animal, such as a mammal or avian
species by administering to the mammal or avian species a
composition comprising glucan, silica, mineral clay and mannan. In
some embodiments the composition comprises between 15% and 40%
silica, between 50% and 81% mineral clay, between 1.0% and 5.0%
.beta.-glucans, and between 1% and 8.0% mannan. In a particular
embodiment the composition consists essentially of .beta.-glucans,
.beta.-1,3 (4)-endoglucanohydrolase, diatomaceous earth, a mineral
clay, and glucomannan.
[0008] In any or all of the above embodiments, the composition can
be administered prophylactically, or when the animal has or is at
risk of developing heat stress. Heat stress may occur when the
animal is exposed to a temperature humidity index of 68 or greater,
75 or greater, or .gtoreq.79.
[0009] In any or all of the above embodiments, heat stress may be
evidenced by a heat stress indicator. Heat stress indicators
include, but are not limited to, feed intake, water consumption,
respiration rate, rectal temperature, milk yield, milk fat, milk
protein, or any combination thereof. In some embodiments the
composition is administered prior to the animal experiencing heat
stress while the animal is experiencing heat stress, and/or after
the animal experiences heat stress. In any or all of the above
embodiments, the animal may be immunosuppressed prior to
administration of the composition, for example as a result of heat
stress.
[0010] In any or all of the above embodiments, the composition may
be administered at set intervals to the animal for an effective
period of time to promote reduction of heat stress. The set
interval can be daily, or it can be more or less frequently than
that. In some embodiments the effective period of time is from 0
days to 200 days, from 1 day to 200 days, from 1 day to 90 days,
from 1 day to 60 days, from 1 day to 45 days, from 1 day to 30
days, from 3 days to 21 days, at least 1 day, at least 3 days, at
least 7 days, at least 30 days, at least 60 days, or at least 90
days.
[0011] In any or all of the above embodiments, the amount
administered to the animal can range from 1 mg/kg body weight per
day to 20 g/kg body weight per day. In any or all of the above
embodiments, the composition may be administered by mixing the
composition with the animal's feed in an amount ranging from 0.1 to
20 kg per ton of feed and providing the composition mixed with the
feed to the animal. In some embodiments, the animal is a bovine and
the amount administered to that animal can range from 0.5 grams to
100 grams daily.
[0012] In any or all of the above embodiments, an amount of the
composition administered to the animal may be increased if the
temperature humidity index is expected to increase or decreased if
the temperature humidity index is expected to decrease.
[0013] In any or all of the above embodiments, the animal may be
administered an amount of the composition effective to alter a heat
stress indicator compared to an animal that has not been
administered the composition. In some embodiments, the method
further includes measuring the heat stress indicator, and selecting
an amount of the composition administered to the animal based at
least in part on the measurement of the heat stress indicator.
[0014] In particular embodiments the animal is a cow, such as, for
example, a dairy cow. Dairy cows may be administered the
composition before lactation. This can be from 90 days prior to
lactation onset to 1 day prior to lactation onset, preferably from
45 days prior to lactation onset to 10 days prior to lactation
onset. The composition may be administered daily prior to lactation
onset. In any or all of the above embodiments, the dairy cow may be
fed the composition for a period of time to increase milk
production and/or reduce milk fat relative to dairy cattle not fed
the composition.
[0015] The animal that is administered the composition may have
reduced water intake relative to animals not fed the composition.
The animal also may have a reduced respiration rate relative to
animals not fed the composition. In other embodiments, the animal
may have a reduced rectal temperature relative to animal not fed
the composition. In particular disclosed embodiments, the animal
may exhibit increased food intake relative to animal not fed the
composition. Also, the animal may exhibit decreased respiratory
alkalosis, rumen acidosis, metabolic acidosis, and any and all
combinations thereof, relative to animals not fed the composition.
In other embodiments, the animal may have decreased serum cortisol
levels relative to animals not fed the composition.
[0016] In any or all of the above embodiments, the method may
further comprise administering to the mammal or avian species a
therapeutic process and/or a therapeutic agent suitable for
treating heat stress. Exemplary therapeutic processes and agents
include, but are not limited to, provision of shade to the animal,
use of a water sprinkler to externally administer water to the
animal, use of a fan to provide air movement, addition of bypass
fats to feed for the animal, meloxicam, a corticosteroid, a
composition comprising one or more electrolytes, an alkalinizing
agent, a direct-fed microbial, or a combination thereof.
[0017] In any or all of the above embodiments, the method may
include administering a first amount of the composition to a group
of animals that have or are at risk of developing heat-induced
stress for a first period of time, measuring a heat stress
indicator of at least one animal of the group of animals, and
administering an adjusted amount of the composition to the group of
animals for a subsequent period of tie, wherein the adjusted amount
is based at least in part on the measurement of the heat stress
indicator.
[0018] In any or all of the above embodiments, the method may
include evaluating at least one heat stress indicator of individual
animals in a group of animals, determining whether one or more of
the individual animals are experiencing heat stress, and
administering the composition to the one or more individual animals
that are experiencing heat stress.
[0019] An exemplary embodiment comprises selecting a dairy cow that
has or is at risk of developing heat induced stress and
administering to the dairy cow before, during, and/or after
lactation a composition comprising glucan, silica, mineral clay,
mannan, and optionally an endoglucanohydrolase, wherein the
composition is administered at set intervals for an effective
period of time sufficient to (1) increase milk production relative
to dairy cattle not fed the composition, (2) lower milk fat
relative to dairy cattle not fed the composition, (3) reduce water
intake relative to dairy cattle not fed the composition, (4) reduce
respiration rate relative to dairy cattle not fed the composition,
(5) reduce rectal temperature relative to dairy cattle not fed the
composition, (6) increase food intake relative to dairy cattle not
fed the composition, (7) decrease respiratory alkalosis, rumen
acidosis, and/or metabolic acidosis relative to dairy cattle not
fed the composition, (8) decrease serum cortisol levels relative to
dairy cattle not fed the composition, or (9) any and all
combinations thereof. The method may further comprise administering
to the dairy cow a therapeutic agent suitable for treating heat
stress.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] FIG. 1 is a bar chart illustrating on-dairy milk yield (kg)
by cows that are administered the disclosed composition and those
that are not, including both initial and on-dairy conditions.
[0021] FIG. 2 is a bar chart illustrating milk yield for control
cows and cows that have been administered the disclosed composition
during thermoneutral conditions (TN), heat stress conditions (HS),
and recovery conditions (Recovery).
[0022] FIG. 3 is a graph illustrating mean milk production by day
for control cows and those that have been administered the
disclosed composition.
[0023] FIG. 4 is a graph illustrating feed intake by day for
control cows and those that have been administered the disclosed
composition.
[0024] FIG. 5 is a bar chart illustrating feed intake for control
cows and cows that have been administered the disclosed composition
during thermoneutral conditions, heat stress conditions, and
recovery conditions.
[0025] FIG. 6 is a graph illustrating somatic cell count by day for
control cows and those that have been administered the disclosed
composition.
[0026] FIG. 7 is a bar chart illustrating mean serum cortisol for
control cows and cows that have been administered the disclosed
composition during thermoneutral conditions (TN) and heat stress
conditions (HS), measured after a certain number of days.
[0027] FIG. 8 is a bar chart illustrating mean serum insulin levels
for control cows and cows that have been administered the disclosed
composition during thermoneutral conditions (TN) and heat stress
conditions (HS), measured after a certain number of days.
[0028] FIG. 9 is a bar chart illustrating mean serum glucose levels
for control cows and cows that have been administered the disclosed
composition during thermoneutral conditions (TN) and heat stress
conditions (HS), measured after a certain number of days.
[0029] FIG. 10 is a bar chart illustrating IL8R receptor gene
expression leukocytes in lactating control cows and cows that have
been administered the disclosed composition during thermoneutral
conditions and heat stress conditions, measured after a certain
number of days.
[0030] FIG. 11 is a bar chart illustrating RANTES protein levels in
control cows and cows that have been administered the disclosed
composition prior to heat stress, during acute heat stress, chronic
heat stress, heat stress recovery and long-term recovery.
[0031] FIG. 12 is a bar chart illustrating serum ACTH levels in
control cows and lactating dairy cows that have been administered
the disclosed composition during thermoneutral conditions (TN) and
heat stress conditions (HS), measured after a certain number of
days.
[0032] FIG. 13 is a graph showing rectal temperature of newly
received steers supplemented with the disclosed composition at a
rate of 4 g/cwt (OmniGen) during the receiving period (28 d) or no
OmniGen (Control) during a subcutaneous lipopolysaccharide
challenge.
[0033] FIG. 14 is a graph showing change in rectal temperature of
newly received steers supplemented with the disclosed composition
at a rate of 4 g/cwt (OmniGen) during the receiving period (28 d)
or no OmniGen (Control) during a subcutaneous lipopolysaccharide
challenge.
[0034] FIG. 15 is a graph showing rectal temperature of newly
received steers supplemented with the disclosed composition at a
rate of 4 g/cwt (OmniGen) during the receiving period (28 d) or no
OmniGen (Control) during a lipopolysaccharide challenge.
[0035] FIG. 16 is a graph showing change in rectal temperature of
newly received steers supplemented with the disclosed composition
at a rate of 4 g/cwt (OmniGen) during the receiving period (28 d)
or no OmniGen (Control) during a lipopolysaccharide challenge.
[0036] FIG. 17 is a bar graph showing overall cortisol
concentrations of newly received steers supplemented with the
disclosed composition at a rate of 4 g/cwt during the receiving
period (OMN-28) or no OmniGen (CON) during a lipopolysaccharide
challenge.
[0037] FIG. 18 is a graph showing cortisol concentrations of newly
received steers supplemented with the disclosed composition at a
rate of 4 g/cwt (OmniGen) during the receiving period (28 d) or no
OmniGen (Control) during a lipopolysaccharide challenge.
[0038] FIG. 19 is a series of bar graphs showing treatment, prior,
and post lipopolysaccharide challenge concentrations of blood urea
nitrogen (BUN) for newly received steers supplemented with the
disclosed composition at a rate of 4 g/cwt (OmniGen) during the
receiving period (28 d) or no OmniGen (Control).
[0039] FIG. 20 is a bar graph showing treatment concentrations of
glucose concentrations for newly received steers supplemented with
the disclosed composition at a rate of 4 g/cwt (OmniGen) during the
receiving period (28 d) or no OmniGen (Control) during a
lipopolysaccharide challenge.
[0040] FIG. 21 is a graph showing glucose concentrations of newly
received steers supplemented with the disclosed composition at a
rate of 4 g/cwt (OmniGen) during the receiving period (28 d) or no
OmniGen (Control) during a lipopolysaccharide challenge.
[0041] FIG. 22 is a series of bar graphs showing treatment, prior,
and post lipopolysaccharide challenge concentrations of
non-esterified fatty acids of newly received steers supplemented
with the disclosed composition at a rate of 4 g/cwt (OmniGen)
during the receiving period (28 d) or no OmniGen (Control).
[0042] FIG. 23 is a series of bar graphs showing treatment, prior,
and post lipopolysaccharide challenge concentrations of
interferon-.gamma. concentrations of newly received steers
supplemented with the disclosed composition at a rate of 4 g/cwt
(OmniGen) during the receiving period (28 d) or no OmniGen
(Control).
[0043] FIG. 24 is two bar graphs showing treatment and post
lipopolysaccharide challenge concentrations of tumor
necrosis-.alpha. (TNF-.alpha.) overall and post LPS challenge
concentrations of newly received steers supplemented with the
disclosed composition at a rate of 4 g/cwt (OmniGen) during the
receiving period (28 d) or no OmniGen (Control).
[0044] FIG. 25 is a graph showing tumor necrosis-.alpha.
(TNF-.alpha.) concentrations of newly received steers supplemented
with the disclosed composition at a rate of 4 g/cwt during the
receiving period (28 d) or no OmniGen (Control) during a
lipopolysaccharide challenge.
[0045] FIG. 26 is a graph showing interleukin-6 (IL-6)
concentrations of newly received steers supplemented with the
disclosed composition at a rate of 4 g/cwt (OmniGen) during the
receiving period (28 d) or no OmniGen (Control) during a
lipopolysaccharide challenge.
[0046] FIG. 27 is a graph showing lymphocyte concentrations of
newly received steers supplemented with the disclosed composition
at a rate of 4 g/cwt (OmniGen) during the receiving period (28 d)
or no OmniGen (Control) during a lipopolysaccharide challenge.
[0047] FIG. 28 is a graph showing the neutrophil to lymphocyte
ratio of newly received steers supplemented with the disclosed
composition at a rate of 4 g/cwt (OmniGen) during the receiving
period (28 d) or no OmniGen (Control) during a lipopolysaccharide
challenge.
DETAILED DESCRIPTION
I. Definitions
[0048] The following explanations of terms and abbreviations are
provided to better describe the present disclosure and to guide
those of ordinary skill in the art in the practice of the present
disclosure. As used herein, "comprising" means "including" and the
singular forms "a" or "an" or "the" include plural references
unless the context clearly dictates otherwise. The term "or" refers
to a single element of stated alternative elements or a combination
of two or more elements, unless the context clearly indicates
otherwise.
[0049] Unless explained otherwise, all technical and scientific
terms used herein have the same meaning as commonly understood to
one of ordinary skill in the art to which this disclosure belongs.
Although methods and materials similar or equivalent to those
described herein can be used in the practice or testing of the
present disclosure, suitable methods and materials are described
below. The materials, methods, and examples are illustrative only
and not intended to be limiting. Other features of the disclosure
are apparent from the following detailed description and the
claims.
[0050] Unless otherwise indicated, all numbers expressing
quantities of components, molecular weights, percentages,
temperatures, times, and so forth, as used in the specification or
claims are to be understood as being modified by the term "about."
Accordingly, unless otherwise indicated, implicitly or explicitly,
the numerical parameters set forth are approximations that may
depend on the desired properties sought and/or limits of detection
under standard test conditions/methods. When directly and
explicitly distinguishing embodiments from discussed prior art, the
embodiment numbers are not approximates unless the word "about" is
recited.
[0051] Animal: This term includes species that are produced for
human consumption or that are domesticated animals. Exemplary
species of such animals are provided herein.
[0052] Administering: Administration by any route to the subject.
As used herein, administration typically refers to oral
administration.
[0053] Mannans: A class of polysaccharides including the sugar
mannose. The mannan family includes pure mannans (i.e., the polymer
backbone consists of mannose monomers), glucomannans (the polymer
backbone comprises mannose and glucose), and galactomannans
(mannans or glucomannans in which single galactose residues are
linked to the polymer backbone). Mannans are found in cell walls of
some plant species and yeasts.
[0054] Mineral Clay: The term "mineral clay" refers to hydrous
aluminum silicates. Mineral clays usually include minor amounts of
impurities, such as potassium, sodium, calcium, magnesium, and/or
iron. Mineral clays typically have a two-layer sheet structure
including tetrahedral silicate sheets and octahedral hydroxide
sheets or a three-layer structure including a hydroxide sheet
between two silicate sheets.
[0055] Therapeutic agent: An agent that is capable of providing a
therapeutic effect, e.g., preventing a disorder, inhibiting a
disorder, such as by arresting the development of the disorder or
its clinical symptoms, or relieving a disorder by causing
regression of the disorder or its clinical symptoms.
[0056] Therapeutically effective amount: A quantity or
concentration of a specified compound or composition sufficient to
achieve a desired effect in a subject being treated for a disorder.
The therapeutically effective amount may depend at least in part on
the species of animal being treated, the size of the animal, and/or
the severity of the disorder.
[0057] Additional disclosure is found in U.S. patent application
Ser. No. 13/566,433, U.S. patent application Ser. No. 13/872,935,
U.S. Patent Publication No. 2013/0017211, U.S. Patent Publication
No. 2012/0156248, U.S. Patent Publication No. 2007/0253983, U.S.
Patent Publication No. 2007/0202092, U.S. Patent Publication No.
20070238120, U.S. Patent Publication No. 2006/0239992, U.S. Patent
Publication No. 2005/0220846, U.S. Patent Publication No.
2005/0180964, and Australian Patent Application No. 2011201420, all
of which are incorporated herein by reference.
II. Composition
[0058] The present disclosure is based on the novel discovery that
a combination of glucan, silica, mineral clay, mannans can
effectively prevent or reduce heat stress in animals. Embodiments
of the disclosed composition comprise one or more components as
disclosed herein. The composition typically comprises glucan (e.g.,
.beta.-1,3 (4)glucan), silica, mineral clay and mannans. In some
embodiments, the composition further comprises an
endoglucanohydrolase, such as .beta.-1,3 (4)-endoglucanohydrolase.
Suitable sources of silica include, but are not limited to, sand,
quartz, diatomaceous earth, and synthetic silica. In certain
embodiments, the mannans comprise glucomannan.
[0059] The components of the composition are prepared by methods
commonly known in the art and can be obtained from commercial
sources. At least some components of the composition (e.g., silica,
mineral clay) also may be present in the environment. .beta.-1,3
(4)-endoglucanohydrolase may be produced from submerged
fermentation of a strain of Trichoderma longibrachiatum.
Diatomaceous earth is available as a commercially-available
acid-washed, product with 95% silica (SiO.sub.2) and with its
remaining components not assayed but consisting primarily of ash
(minerals) as defined by the Association of Analytical Chemists
(AOAC, 2002). The mineral clays (e.g., aluminosilicates) used in
this composition may be any of a variety of commercially-available
clays including, but not limited to, montmorillonite clay,
bentonite and zeolite. Glucan and mannans can be obtained from
plant cell walls, yeast (e.g., Saccharomyces cerevisiae, Candida
utilis), certain fungi (e.g., mushrooms), and bacteria.
[0060] In one embodiment, the composition includes 1-40 wt %
silica, 1-25 wt % glucan and mannans, and 40-92 wt % mineral clay.
In another embodiment, the composition comprises 5-40 wt % silica,
2-15 wt % glucan and mannans, and 40-80 wt % mineral clay. In
another embodiment, the composition comprises 20-40 wt % silica,
4-10 wt % glucan and mannans, and 50-70 wt % mineral clay. In
another embodiment, the composition comprises 15-40 wt % silica,
1-15 wt % glucans, 0-10 wt % mannans, and 50-81 wt % mineral clay.
In another embodiment, the composition comprises 15-40 wt % silica,
1.0-5.0 wt % glucans, 1.0-8.0 wt % mannans, and 50-81 wt % mineral
clay. In another embodiment, the composition comprises 20-30 wt %
silica, 1.0-3.5 wt % glucans, 1.0-6.0 wt % mannans, and 60-75 wt %
mineral clay.
[0061] In some embodiments, .beta.-glucans and mannans are obtained
from yeast cell wall extract, and Composition I comprises 1-40 wt %
silica, 1-30 wt % yeast cell wall extract, 40-92 wt % mineral clay.
In one embodiment, Composition I comprises 10-40 wt % silica, 5-20
wt % yeast cell wall extract, and 40-80 wt % mineral clay. In
another embodiment, Composition I comprises 15-30 wt % silica, 5-15
wt % yeast cell wall extract, and 55-70 wt % mineral clay.
[0062] In any of the above embodiments, the composition may further
comprise an endoglucanohydrolase, such as .beta.-1,3
(4)-endoglucanohydrolase. The composition may include from at least
0.05 wt % endoglucanohydrolase to 5 wt % endoglucanohydrolase, such
as from 0.05-3 wt % .beta.-1,3 (4)-endoglucanohydrolase. In one
embodiment, Composition I consists essentially of 0.1-3 wt %
.beta.-1,3 (4)-endoglucanohydrolase, 20-40 wt % silica, 2-20 wt %
glucan and mannans, and 50-70 wt % mineral clay. In another
embodiment, the composition consists essentially of 0.2-3 wt %,
.beta.-1,3 (4)-endoglucanohydrolase, 20-40 wt % silica, 4-10 wt %
glucan and mannans, and 50-70 wt % mineral clay. In any of the
above embodiments, the silica may be provided by diatomaceous
earth. In any of the above embodiments, the glucans may be
.beta.-glucans. In some embodiments, the .beta.-glucans can be
obtained from yeast, or other materials, such as fungi, algae, or
the like. In any of the above embodiments, the mannans may comprise
glucomannan.
[0063] The glucan and mannans (or yeast cell wall extract) can be
prepared by a method commonly known in the art. In an independent
embodiment, it can be a commercial source of .beta.-1,3 (4) glucan
and glucomannan derived from primary inactivated yeast
(Saccharomyces cerevisiae) with the following chemical
composition:
[0064] Moisture 3.5-6.5%
[0065] Proteins 1-6%
[0066] Fats 0-0.5%
[0067] Phosphorus 0-0.2%
[0068] Mannans 9-20%
[0069] .beta.-1, 3-(4) glucan 9-18%
[0070] Ash 75-85%; and in some embodiments, the composition can
further comprise Dry matter 97-98%.
[0071] In another independent embodiment, the composition can
comprise:
[0072] Moisture 2-3%
[0073] Dry matter 97-98%
[0074] Proteins 14-17%
[0075] Fats 20-22%
[0076] Phosphorus 1-2%
[0077] Mannans 22-24%
[0078] .beta.-1, 3-(4) glucan 24-26%
[0079] Ash 3-5%.
[0080] The mineral clays (aluminosilicates) used in this
composition may be fulfilled by any of a variety of
commercially-available clays including, but not limited to,
montmorillonite clay, bentonite and zeolite.
[0081] In an independent embodiment of the composition, silica,
glucan and mannans, and mineral clay are combined at 1-40%, 1-25%
and 40-92%, respectively. In an independent embodiment of the
composition, .beta.-1,3 (4)-endoglucanohydrolase, diatomaceous
earth, yeast cell wall extract and mineral clay are combined at
0.05-3%, 1-40%, 1-20% and 40-92%, respectively. In an independent
composition, .beta.-1,3 (4)-endoglucanohydrolase, diatomaceous
earth, yeast cell wall extract and mineral clay are combined at
0.1-3%, 5-40%, 2-10% and 40-80%, respectively. In another
independent embodiment of the composition, .beta.-1,3
(4)-endoglucanohydrolase, diatomaceous earth, yeast cell wall
extract and mineral clay are combined at 0.2-3%, 30-40%, 4-6% and
50-65%, respectively.
[0082] In some embodiments, the composition includes additional
components. Additional components may be used for any desired
purpose, such as a substantially biologically inert material added,
for example, as a filler, or to provide a desired beneficial
effect. For example, the composition may include a carbonate
(including a metal carbonate such as calcium carbonate), kelp, a
vitamin (such as a niacin supplement or vitamin B-12 supplement),
biotin, d-calcium pantothenate, choline chloride, thiamine
mononitrate, pyridoxine hydrochloride, menadione
dimethylpyrimidinol bisulfite, riboflavin-5-phosphate, folic acid,
soybean oil, calcium aluminosilicate, rice hulls, mineral oil, or
any combination thereof.
[0083] The composition may be formulated in any suitable form,
including a powder, a granule, a pellet, a solution, or a
suspension. In one embodiment, the composition can be a dry,
free-flowing powder suitable for direct inclusion into a
commercially-available feed, food product or as a supplement to a
total mixed ration or diet. The powder may be mixed with either
solid or liquid feed or with water. In another embodiment, the
composition can be formed into pellets.
[0084] In one embodiment, when incorporated directly into feeds,
the composition may be added in amounts ranging from 0.1 to 100 kg
per ton, such as from 0.1 to 20 kg per ton (2000 pounds) of feed.
In some embodiments, the composition can be added to animal
feedstuffs or to food in amounts from 0.1 kg to 50 kg per ton, from
0.1 to 20 kg per ton, or from 0.5 kg to 10 kg per ton of feed. In
certain embodiments, the composition may be added to feeds in
amounts ranging from 1 to 5 kg per ton of feed.
[0085] When expressed as a percentage of dry matter of feed, the
composition may be added to animal feedstuffs or to foods in
amounts ranging from 0.01 to 2.5% by weight, such as from 0.0125%
to 2% by weight. In one embodiment, the composition can be added to
animal feedstuffs or to food in amounts from 0.05 to 1.5% by
weight, such as from 0.06% to 1% by weight. In another embodiment,
the composition can be added in amounts from 0.1 to 0.7% by weight,
such as from 0.125% to 0.5% by weight of feed.
[0086] Alternatively, the composition may be fed directly to the
animal as a supplement in amounts of from 0.01 gram to 20 gram per
kilogram of live body weight per day, such as from 0.01 gram to 10
gram per kilogram, 0.01 gram to 5 gram, 0.01 gram to 1 gram, 0.015
gram to 1 gram, or 0.02 gram to 0.4 gram per kilogram of live body
weight per day. In some embodiments, the composition may be
provided for use with many species in amounts of from 0.05 grams to
0.20 grams per kilogram of live body weight per day.
[0087] Additionally, the composition may be fed to mammalian
animals or avian species as a supplement (in combination with other
feed, or alone) in amounts ranging from 10 grams per head per day
to 70 grams per head per day, such as from 40 grams per head per
day to 70 grams per head per day, from 45 grams per head per day to
70 grams per head per day, or from 50 grams per head per day to 70
grams per head per day. In exemplary embodiments, the composition
can be provided to bovines in an amount ranging from 50 grams per
head per day to 60 grams per head per day, such as 56 grams per
head per day.
[0088] The physical form of the composition can be a dry,
free-flowing powder which is suitable for direct inclusion into a
feed, food product or as a supplement to a total mixed ration or
diet.
[0089] Alternatively, the composition contained in the present
composition may be fed directly to mammalian or avian species as a
supplement in amounts 0.016 grams/kg to 0.37 grams/kg of live body
weight per day. In an independent embodiment, the composition may
be provided to mammalian and avian species in amounts of 0.10
grams/kg to 0.20 grams/kg of body weight per day.
[0090] In particular disclosed embodiments, the composition may be
administered to the mammal or avian species using a carrier. The
carrier may be any carrier known to a person of ordinary skill in
the art as being suitable for combining with a feed composition,
such as molasses. In some embodiments, the composition includes
additional ingredients. For example, the composition includes
calcium carbonate, dried kelp, niacin supplement, biotin, d-calcium
pantothenate, vitamin B-12 supplement, choline chloride, thiamine
mononitrate, pyridoxine hydrochloride, silicon dioxide,
riboflavin-5-phosphate, folic acid, soybean oil, or any combination
thereof.
[0091] One of skill in the art can appreciate that the amount of
the claimed composition fed can vary depending upon the animal
species, size of the animal and type of the feedstuff to which the
claimed composition is added.
III. Reducing Heat Stress
[0092] In particular disclosed embodiments, the composition
disclosed herein may be used to prevent and/or promote reduction of
heat stress in animals. Heat stress can impair production,
reproduction, and animal health. For example, heat stress can lead
to increased respiratory rate, increased body temperature,
increased fluid intake, decreased feed intake, decreased weight
gain, decreased milk yield, decreased reproduction, respiratory
alkalosis, ruminal alkalosis, metabolic acidosis, increased
oxidative stress, decreased immune function, and/or increased cell
damage and death. Accordingly, any of the foregoing factors may be
assessed as heat stress indicators. In some embodiments, the heat
stress indicator is feed intake, water consumption, respiration
rate, rectal temperature, milk yield, milk fat, milk protein, an
immune biomarker, or any combination thereof. In certain
embodiments, the heat stress indicator is feed intake, water
consumption, respiration rate, rectal temperature, milk yield, milk
fat, milk protein, or any combination thereof.
[0093] In some embodiments, heat stress conditions occur when a
temperature humidity index (THI) is greater than 68, such as
greater than 75, .gtoreq.79 (regarded as a dangerous level), or
.gtoreq.84 (regarding as an emergency level). In another
embodiment, heat stress conditions occur when the heat index,
commonly reported by the media for humans, is above 100, such as
above 110, above 115, or above 120.
[0094] The composition may be administered to an animal that is
susceptible to heat stress or that suffers from heat stress. In one
embodiment, the composition is administered to the animal when the
temperature humidity index is, or is expected to be, greater than
68. The composition may be administered to the animal at set
intervals. For example, the composition may be administered to the
animal on a daily basis. In some embodiments, a daily amount of the
composition is divided and administered to the animal at two or
more feedings.
[0095] In some embodiments, the animal can be an animal raised for
human consumption including, but are not limited to mammals, such
as livestock (e.g., feed or dairy cattle) or pigs; avians, such as
domestic fowl (e.g., chicken, turkey, goose, duck, cornish game
hen, quail, pheasant, guinea-fowl, ostrich, emu, swan, or pigeon);
or fish (e.g., salmon, trout and the like). In other embodiments,
the animal can be a domestic animal, such as a dog, cat, fish, or
rabbit. In some other embodiments, the animal can be a ruminant
species, such as a sheep, goat, cow, deer, bison, buffalo, or
llama. In yet other embodiments, the animal can be an ungulate,
such as a horse, donkey, or pig.
[0096] The disclosed method and composition can be used to promote
reduction of heat stress in any mammalian (including human), avian,
or aquatic species. In a particular embodiment, the compositions
are administered to livestock animals, including both ruminants
(e.g., cattle, sheep, goats, cows, deer, bison, buffalo) and
non-ruminants (e.g., pigs, horses, sows).
[0097] The composition disclosed herein may be administered in an
amount effective to promote reduction of heat stress in animals,
such as dairy cows, particularly lactating dairy cows. The
composition may be administered pre- or post-calving for a suitable
number of days. The composition also may be administered prior to
lactation or after lactation. For example, the composition may be
administered to the animal for a period of from 1 day prior to
lactation onset to 100 days prior to lactation onset. In other
disclosed embodiments, the composition may be administered to the
animal for 40 days to 100 days post calving, or for 45 days to 95
days post calving, or for 50 days to 90 days post calving.
[0098] In some embodiments, a dairy cow is fed the composition for
a period of time to increase milk production relative to dairy
cattle not fed the composition. In particular embodiments, the
dairy cow has lower milk fat relative to dairy cattle not fed the
composition. In one embodiment, the dairy cow had reduced water
intake relative to dairy cattle not fed the composition. In another
embodiment, the dairy cow had a reduced respiration rate relative
to dairy cattle not fed the composition. In yet another embodiment,
the dairy cow had a reduced rectal temperature relative to dairy
cattle not fed the composition. The dairy cow may have increased
food intake relative to dairy cattle not fed the composition. In
particular disclosed embodiments, the dairy cow may have decreased
respiratory alkalosis, rumen acidosis, metabolic acidosis, and any
and all combinations thereof, relative to dairy cattle not fed the
composition. In another embodiment, the dairy cow has decreased
serum cortisol levels relative to dairy cattle not fed the
composition.
[0099] In additional disclosed embodiments, the composition may be
administered prophylactically. In certain embodiments, the
composition may be administered prophylactically and continuously.
For example, the animal may be administered the composition every
day for a certain period of time prior to and during lactation.
[0100] The disclosed composition may be formulated for
administration to animals in order to prevent and/or promote
reduction of heat stress. In particular disclosed embodiments, the
effective amount of the composition administered can be
affirmatively chosen based on the ability of that effective amount
to promote heat stress reduction in an animal based on particular
factors disclosed herein. In particular disclosed embodiments, the
effective amount may be determined by administering a particular
first dose to the animal, monitoring the animal during heat stress,
and then adjusting the dose in order to determine an effective
amount of a second dose that will ameliorate, or further ameliorate
the heat stress of the animal.
[0101] In one embodiment, the composition is administered to a
group of animals that have or are at risk of developing heat
stress. The composition may be administered to the group of animals
in a first amount for a first period of time. Subsequently a heat
stress indicator of at least one animal in the group of animals is
measured. Based at least in part on the heat stress indicator
measurement, the amount of composition administered to the group of
animals for a subsequent period of time may be adjusted. For
example, the amount may be increased if the heat stress indicator
measurement indicates that the animals are stressed. Alternatively,
the amount may be decreased if the heat stress indicator
measurement indicates that the animals are not stressed or have
relatively little heat stress.
[0102] In another embodiment, individual animals in a group of
animals are evaluated for heat stress. Heat stress can be
evaluated, for example, by measuring a level of one or more heat
stress indicators. Based at least on the measurement, a
determination is made whether one or more of the individual animals
is experiencing heat stress. Individual animals in the group that
are experiencing heat stress may be selected to receive the
composition for a period of time effective to promote reduction of
heat stress.
[0103] In particular disclosed embodiments, the composition may be
administered with a molasses carrier once or multiple times a day
(e.g., from two to five times per day). The composition may be
mixed into the total mixed ration of feed that can be provided to
the mammal or avian species, such as in the top one-fourth, top
one-third, or top one-half of the total mixed ration.
[0104] In some embodiments, the animal is further administered a
therapeutic process and/or a therapeutic agent suitable for
treating stress. Exemplary therapeutic processes and agents
include, but are not limited to, provision of shade to the animal,
use of water sprinklers to externally administer water to the
animal, use of a fan to provide air movement, addition of high-fat
feeds or bypass fats, meloxicam, corticosteroids (isoflupredone,
fludrocortisone, triamcinolone, dexamethasone, betamethasone,
flumethasone, methylprednisolone acetate, methylprednisolone sodium
succinate), oral electrolytes (e.g., sodium, glucose, glycine,
potassium, chloride) alkalinizing agents (e.g., bicarbonate,
acetate, and/or citrate salts), direct-fed microbials, and
combinations thereof.
[0105] The ability of the composition to reduce and/or prevent heat
stress may be determined by comparing heat stress indicators with
animals that are not administered the composition. In particular
disclosed embodiments, heat stress indicators may be used to
determine the effect of the composition on heat stress. Suitable
heat stress indicators/factors include, but are not limited to,
feed intake, milk yield, milk fat, milk protein, water consumption,
respiration rates, rectal temperatures, and combinations
thereof.
[0106] In particular disclosed embodiments, animals that are
administered the composition will have a higher feed intake during
heat stress compared to animals that are not administered the
composition. The feed intake during heat stress may increase from 2
kg to 10 kg (or from 2 kg to 8 kg, or from 2 kg to 6 kg, or 2 kg to
4 kg) compared to the feed intake of an animal of the same species
has not been administered the composition. In exemplary
embodiments, a bovine that is administered the composition will
have feed intake of 3 kg higher than a bovine who has not been
administered the composition.
[0107] The milk yield also may be maintained or increased (as
compared to yields from animals that are not administered the
composition) during heat stress by administering the composition.
For example, the milk yield may increase by 1 kg, 2 kg, 3 kg, 4 kg,
up to 10 kg using the disclosed composition. In particular
disclosed embodiments, animals that are provided the composition
will produce milk having lower milk fat and/or milk protein (in
terms of percentage). For example, milk fat may be reduced from
0.2% to 1%, or from 0.2% to 0.8%, or from 0.2% to 0.6%, with
exemplary embodiments including a 0.4% reduction. In one
embodiment, a dairy cow fed the composition produces a milk fat
percentage of 3.8%, whereas dairy cattle not fed the composition
have a milk fat percentage of 4.2%.
[0108] Water consumption, respiration rates, and serum cortisol
levels also are lowered by administering the disclosed composition,
with some embodiments exhibiting from 4 to 10 fewer respirations
per minute. Another factor indicating that the disclosed
composition is capable of reducing heat stress is rectal
temperature, which typically may be lowered by 0.10.degree. C. to
0.3.degree. C., in comparison to the rectal temperatures taken from
animals that are not administered the composition. Additionally,
animals that have been administered the disclosed composition may
have decreased respiratory alkalosis, rumen acidosis, metabolic
acidosis, and combinations thereof in comparison to animals that
were not administered the composition.
IV. Examples
Example 1
[0109] A total of 60 cows on a commercial dairy in Arizona were
balanced for DIM, parity and milk production and assigned to 1 of 2
treatment groups fed the disclosed composition (OmniGen-AF.RTM.
[OG] comprising between 15% and 40% silica, between 50% and 81%
mineral clay, between 1.0% and 5.0% .beta.-glucans, between 0.05%
and 3.0% .beta.-1,3 (4)-endoglucanohydrolase and between 1% and
8.0% mannan, 30 cows) or control (CON, 30 cows) diets for 52 d post
calving. At 52 d of lactation cows were randomly selected (n=12)
from both groups (6 OG and 6 CON) and housed in environmentally
controlled modules for 21 d. The OG was top-dressed 2.times./d with
molasses as the carrier and the CON cows received the molasses
carrier 2.times./d. Both were mixed into the top one-third of the
TMR (total mixed ration). During the environmental room phase of
the study cows fed OG had higher feed intake than CON during heat
stress (HS) (46.8 kg vs. 42.9 kg, P<0.0001) and no difference
during thermoneutral (TN). A temperature-humidity index (THI)
threshold of 68 or greater was used to achieve HS. Feeding OG
maintained a numerical 1 kg milk yield advantage compared with CON
(30.3 kg vs. 31.4 kg, P=0.26) during HS but not during TN. Cows fed
OG had lower milk fat (%) (4.2% vs. 3.8%, P=0.02) and milk protein
(%) (P=0.04). There was no difference in 3.5% FCM between
treatments. Water consumption was lower (12.41/d in OG treated
cows, P<0.01) than control cows. Respiration rates were lower in
treated cows at 1400 h and 1700 h (4.7 and 8.4 less
respirations/min, P=0.05, <0.001) and rectal temperatures were
also lower (0.15.degree. C. and 0.25.degree. C. lower that CON,
P=0.05, <0.001) in treated cows. Feeding OG reduced
physiological responses to heat stress in lactating dairy cows.
Example 2
[0110] A total of 30 cows on a commercial dairy in Arizona were
balanced for DIM, parity and milk production and assigned to 1 of 2
treatment groups fed the disclosed composition (OmniGen-AF.RTM.
[OG] comprising between 15% and 40% silica, between 50% and 81%
mineral clay, between 1.0% and 5.0% .beta.-glucans, between 0.05%
and 3.0% .beta.-1,3 (4)-endoglucanohydrolase and between 1% and
8.0% mannan, 15 cows) or control (CON, 15 cows) diets for 90 d post
calving. At 90 d of lactation, cows were randomly selected (n=12)
from both groups (6 OG and 6 CON) and housed in environmentally
controlled modules for 21 d at the University of Arizona. The OG
was top-dressed 2.times./d with molasses as the carrier. The CON
cows received the molasses carrier 2.times./d. Both were mixed into
the top one-third of the TMR. During the environmental room phase
of the study, cows fed OG had higher feed intake than CON during
heat stress (HS) (46.8 kg vs. 42.9 kg, P<0.0001) and no
difference during thermoneutral (TN). A temperature-humidity index
(THI) threshold of 68 or greater was used to achieve HS. Feeding OG
maintained a numerical 1 kg milk yield advantage compared with CON
(30.3 kg vs. 31.4 kg, P=0.26) during HS but not during TN. Cows fed
OG had lower milk fat (%) (4.2% vs. 3.8%, P=0.02) and milk protein
(%) (P=0.04). There was no difference in 3.5% FCM between
treatments. Water consumption was lower (12.41/d in OG treated
cows, P<0.01) than control cows. Respiration rates were lower in
treated cows at 1400 h and 1700 h (4.7 and 8.4 less
respirations/min, P=0.05, <0.001) and rectal temperatures were
also lower (0.15.degree. C. and 0.25.degree. C. lower that CON,
P=0.05, <0.001) in treated cows. Feeding OG reduced
physiological responses to heat stress in lactating dairy cows.
[0111] Experimental Design:
[0112] The study consisted of two phases; 1) the commercial dairy,
and 2) the controlled environmental chambers. During the commercial
dairy phase, multiparous lactating Holstein cows (n=30) were
balanced by DIM, milk production and parity (91.+-.5.9 DIM,
36.2.+-.2.5 kg/d, and 3.1.+-.1.4). Cows were separated into one of
two groups. The control group received the base TMR with no
supplement. The treatment group was fed the base diet plus 56
g/head/day of OmniGen-AF.RTM. composition (OG) mixed into the TMR.
Daily milk production was measured. The dairy phase lasted for 45
days. The dairy portion was used to meet the manufacture's
recommended 45 d feeding for OG composition to function.
[0113] After the on-dairy portion was complete, 12 cows (6 control
and 6 treatment) were housed in the environmentally controlled
rooms at the Agricultural Research Center (ARC). Cows continued the
ARC portion in the same treatment groups from the on-dairy
portion.
[0114] The ARC portion lasted for 21 days. Cows were subjected to 7
days of TN conditions, 10 days of HS, and 4 days of recovery (TN).
The diurnal cycle during thermoneutral (TN) and recovery maintained
a temperature humidity index (THI)<68. During HS, the THI was
greater than 68 for 16 hours/day. Temperatures mimicked ambient
temperatures at a southwest United States dairy during summer heat
and TN conditions. Fresh feed was provided twice daily and cows
were individually fed. Control animals received base TMR, and OG
cows received 56 g/head per day, split between two meals. Feed
intake, milk production, and milk composition were measured daily.
Rectal temperatures and respiration rates were recorded 3.times./d
(600, 1400, and 1800 h). Blood samples were taken by venipuncture
from the tail (coccygeal) vein on days 7 (TN), 8 (HS), 10 (HS), 17
(HS) and 18 (TN) during the ARC segment. Samples were collected 6
times per day (0400, 0800, 1200, 1600, 2000, and 2400 h) on days 7,
8, 17, and 18, and once per day on day 14 (0800 h). Blood was
collected in Vacutainer (BD Vacutainer, Franklin Lakes, N.J.) tubes
containing sodium heparin for plasma and in sterile blank tubes for
serum.
[0115] Statistical analyses were performed using the PROC MIXED
procedure (version 9.3, SAS Institute, Cary, N.C.). Cow was the
experimental unit (ARC portion). Data is presented in least square
means with significance declared with a P-value.ltoreq.0.05. (See
Table 1, below).
[0116] There were no initial differences in milk yield
(control=38.6 kg/day and treatment=38.6 kg/day) at the start of the
on-dairy phase of the study. There was a numerical advantage to
feeding OG (FIG. 1) of 1.5 kg of milk/day, but this was not
significant (control=36.8 kg/day and treatment=38.3 kg/day).
[0117] There was a period effect on milk yield (P<0.01) during
the environmental room (ARC) phase associated with a decline in
milk yield in both groups during HS. Milk yield at the ARC
(P<0.23) did not differ between control and OG fed groups (FIGS.
2, 3), however, there was a numerical advantage (1.1 kg/day) for
cows fed OG during HS (P<0.26) which was similar to the pattern
in milk yield noted during the on-farm phase
[0118] Feeding the disclosed composition to heat stressed dairy
cows maintained feed intake during heat stress. Feed intakes in the
two groups did not differ during TN but was higher during HS in
OG-fed cows (46.8 kg/d and 42.9 kg/d, P<0.01, FIGS. 4, 5; Table
1).
[0119] Milk protein (%) and fat (%) were lower in OG-fed cows
(Table 1) during HS but not during TN. There was no difference in
FCM or protein yield between treatments. Cows fed OG displayed
decreased somatic cell count (SCC) compared to control cows (59.4
and 26.3.times.1000, P<0.03; Table 1) with the greatest
difference during the recovery period (FIG. 6). There was a spike
in SCC around day 5 (TN) and during recovery around day 17 (FIG.
6).
TABLE-US-00001 TABLE 1 Effects of OmniGen-AF supplementation in
heat-stressed lactating dairy cows Control OmniGen-AF Item TN HS
Recovery TN HS Recovery SEM P-value Feed intake (kg) 46.1 42.9 47.5
47.1 46.8* 49.1 1.04 0.01 Milk yield (kg) 33.1 30.3 30.4 33.9 31.4
31.3 1.02 0.23 Fat (%) 4.03 4.22 4.16 3.94 3.82 3.83 0.22 0.04 FCM
(kg/d) 35.0 33.7 33.7 34.7 32.8 32.6 1.45 0.39 Protein (%) 2.95
2.98* 2.86 2.95 2.86 2.79 0.07 0.15 Protein (kg) 0.98 0.89 0.90
1.00 0.93 0.92 0.30 0.13 Lactose (%) 4.87 4.85 4.99 4.89 4.78 4.96
0.08 0.61 SCC 20.3 23.9 59.4* 19.6 22.9 26.3 9.12 0.03 *= P-value
.ltoreq.0.05 and indicates the higher value
[0120] Respiration rate and rectal temperatures did not differ
between treatments during TN; however, during HS, OG reduced
respiration rate, (Table 2, P<0.01) in both environments at 1400
h and in HS animals at 1800 h when environmental heat load was
greatest. Rectal temperatures were lower in cows fed OG at 1400 and
1800 h compared to controls when environmental heat loads were
maximal.
TABLE-US-00002 TABLE 2 Effects of OmniGen-AF supplementation and
environment on respiration rate and rectal temperature in lactating
dairy cows Control OmniGen-AF P- Item TN HS Recovery TN HS Recovery
SEM value Resp/min 600 26.9 31.9 28.3 26.6 30.4 27.9 1.40 0.40 1400
34.3 63.1* 35.3 30.1 58.3 35.5 2.99 0.20 1800 34.9* 60.8* 32.1 29.5
52.4 29.7 2.62 0.01 Rectal Temp (.degree. C.) 600 38.2 38.0 37.9
38.2 38.1 38.1 0.05 0.26 1400 38.0 38.7* 38.0 38.1 38.5 38.1 0.09
0.77 1800 38.2 39.1* 38.2 38.2 38.8 38.3 0.08 0.25 *P .ltoreq. 0.05
and indicates the higher value.
[0121] Hormones in plasma are important as potential indicators of
the physiological status of a cow and reflect the physiological
compensations a cow undergoes at various stages of lactation and
exposure to HS. Serum cortisol levels were highest on day 8 (first
day of HS, FIG. 7). This is in agreement with prior reports that
acute but not chronic HS is associated with increases in
circulating cortisol concentrations (Christian and Johnson, 1972,
Wise et al., 1988). OG treated cows had significantly lower serum
cortisol on day 8 (0.8372 vs. 0.4838 .mu.g/dL for control and OG
respectively, P<0.006) and did not differ on other days. This
suggests that OmniGen may reduce impact of acute stress on the
cortisol response in lactating dairy cows.
[0122] Serum insulin and plasma glucose levels (FIGS. 8 and 9) were
not different between groups (P=0.8248 and 0.945). Serum insulin
concentrations in both groups rose during the latter part of the HS
period and during the recovery period. The reason for this pattern
is unknown.
[0123] The immune function of cattle on this study was evaluated by
looking at the expression of the interleukin-8 receptor (FIG. 10)
and expression of Regulated on Activation, Normal T Expressed and
Secreted (RANTES) protein (FIG. 11) which is a member of the
interleukin-8 family of cytokines.
[0124] Heat stress exposure was mild to moderate in this study. The
threshold for heat stress in lactating dairy cows is a THI>68,
respiration rates>60 bpm, and rectal
temperatures>38.5.degree. C. (Zimbleman et al., 2009). OG
reduced impact of thermal stress on stress of lactating dairy cows.
Cows fed OG had reduced rectal temperatures and respiration rates
during periods of peak thermal load. Respiration rates in treated
cows did not exceed 60 bpm and mean rectal temperatures were 0.2 to
0.3.degree. C. cooler. OG fed cows displayed higher feed intakes
during HS as well. Cows fed OG also displayed a lower cortisol
spike on the first day of heat stress.
[0125] Milk yield decreased with heat stress in both control
animals and the OG fed animals. However, feed intake was unchanged
in cows fed OG and milk yields were numerically higher. Changes in
SCC were consistent between groups. Cows fed OG displayed decreased
SCC compared to control cows with the greatest difference during
the recovery period.
[0126] Serum cortisol levels were similar to previous findings
(Christison and Johnson, 1972) and increased within the first day
of heat exposure. The animals in the ARC had higher cortisol levels
compared to published levels, but the confinement and changes in
surrounding from the dairy to the ARC may account for some of the
changes.
[0127] Cytokine (RANTES) gene expression was higher in cows fed OG
during the HS portion of the study but not during recovery. The
elevated cytokine gene expression may be associated with improved
immune function in cows fed OmniGen-AF.
Example 3
[0128] Embodiments of the disclosed composition, such as
OmniGen-AF.RTM. composition comprising between 15% and 40% silica,
between 50% and 81% mineral clay, between 1.0% and 5.0%
.beta.-glucans, between 0.05% and 3.0% .beta.-1,3
(4)-endoglucanohydrolase and between 1% and 8.0% mannan (OG), have
been demonstrated to reduce physiological measures of heat stress
(e.g., body temperature, respiration rate, and water intake) in
cattle subjected to temperature and humidity conditions above their
thermoneutral zone. Concomitantly, in OG-fed animals, feed intake
was significantly increased and milk yield was numerically
increased during periods of thermal stress. Furthermore, measures
of immune function were improved along with reduced cortisol
concentrations during acute heat stress. Paradoxically,
adrenocorticotrophic hormone (ACTH) which regulates cortisol
secretion from the adrenal cortex was significantly increased in
animals fed OG (FIG. 12). This suggests that either the adrenal is
less responsive to ACTH or that corticoid binding globulin is
increased in animals fed OG. Both conditions would result in
increased ACTH secretion from the pituitary since negative feedback
in both cases would be reduced. The objective of this study is to
determine if feeding OG to lactating dairy cows decreases adrenal
responsiveness to ACTH or increases corticosteroid binding globulin
in either thermoneutral or heat stress conditions.
[0129] Heat stress in dairy cows may result in reductions in dry
matter intakes and milk yields and elevated somatic cell counts,
respiration rates, body temperature and plasma heat shock protein
and serum cortisol. Feeding OG may reduce the impact of heat stress
on these measures of stress response. Feeding OG to non-heat
stressed lactating dairy cows for 52 days prior to a heat stress
bout may increase the expression of immune markers of neutrophil
function (L-selectin, IL8-R and IL-1B) associated with altered
secretion of cortisol from the adrenal gland. This may be
associated with reduced cortisol secretion in response to ACTH
infusion in cows fed OG. Heat stress may reduce the expression of
immune markers of neutrophils even in cows supplemented with OG.
However, the reduced cortisol secretion response may also be
present in heat stressed dairy cows fed OG.
[0130] Experimental Design:
[0131] Animal Selection and Treatment Assignment:
[0132] Thirty lactating dairy cows will be assigned to one of two
treatments (15 head/treatment). Treatment 1 cows will be fed a
control diet without OG, and Treatment 2 cows will be fed the
control diet plus OG. OG will be pre-blended in a grain mix to
provide 56 g/h/d. Each cow will be housed in an individual
tie-stall where individual feed and water intake can be controlled
and recorded prior to moving to the Agricultural Research Center
(ARC). After 45 days on study, all cows will be given a low dose
ACTH challenge (20 mg) via tail vein infusion, and blood samples
will be taken at time zero, 1 hour, 4 hours and 8 hours after ACTH
challenge. Subsequently, a subgroup of 12 cows, which have been on
their respective diets on a commercial dairy for 55 days prior to
the moving to the ARC environmental controlled rooms, will be
added. From the 30 original cows assigned to treatments, 6 cows
from each treatment will be selected for the trial and will
continue on their respective treatment diets while in the ARC. Cow
will be the experimental unit.
[0133] Blood Sample Schedule:
[0134] Blood samples will be collected on all treatment cows while
at the commercial dairy at days 1, 21 and 42 after initial
treatment assignment. On days 45-50 ten cows each day will be
subjected to low dose ACTH challenge. On each sampling day,
approximately 2 hours after the morning feeding, cows will be
restrained in headlocks, taking care to avoid stressing the
animals; tail vein blood samples will be obtained from each animal
and then infused via tail vein or tail artery with 20 .mu.g (=2IU)
of a synthetic analogue of ACTH (ACTH1-24, Synacthen.RTM.--Novartis
Pharma AG--Stein, CH). Blood samples will also be taken 30 and 60
minutes after ACTH injection, leaving cows restrained in the head
locks. All the blood samples will be collected in vacuum Li-heparin
tubes, and immediately stored in iced water. In the blood samples,
packed cell volume (PCV) will be determined and, after
centrifugation (3500 g for 16 min. at 6.degree. C.), plasma
cortisol will be measured by the RIA method (Coat-A-Count; DPC, Los
Angeles, Calif., USA). The integrated response of cortisol over 60
min will be evaluated as area under the curve. For the statistical
evaluation, data will be subjected to ANOVA using GLM procedure
(SAS Inst. Inc., Cary, N.C., release 9.1) including in the model
cow, dietary treatment (control or OG) stage of lactation as main
factors. A repeated measures analysis will also be conducted on the
challenge data.
[0135] At arrival to the ARC, cows will be weighed, fitted with
halters and blood samples collected. Cows will remain in the ARC
chambers for 21 days, the first 7 days at thermal neutral (TN)
conditions followed by 10 days of heat stress (HS) and then 4 days
in TN conditions. On d 7 of TN, d 8 (HS), d 17 (HS), d 18 (TN) and
d 21 (TN) cows will be bled in 4 h intervals at 0400, 0800. 1200,
1600, 2000, and 2400 h following a low dose (20 mg) of ACTH infused
via the tail vein.
[0136] Physiological/Behavior Metrics:
[0137] To assess the effectiveness of the imposed heat stress
model, known physiological and behavioral responses will be
measured and recorded daily. These will include feed intake, water
consumption, milk yield, milk somatic cell concentrations
(cells/ml), respiration rate, rectal temperature and skin
temperature measurements (3.times./d at 0600, 1400 and 1800 h).
Milk samples will be collected and stored in vials containing
bronopol tablets for preservation and stored at 4.degree. C. until
analysis. Analysis will be done by infrared analysis. All treatment
and health events will be recorded daily.
[0138] Blood collected for immune biomarkers will be collected and
preserved according to protocol (Wang, et al., 2003). The immune
biomarkers neutrophil L-selectin, IL8-R and IL-1B will be evaluated
on each sample. In addition, on days 17 and 18 from the 0800 and
1600 h samples, neutrophils will be purified and assessed for
RANTES (regulated on activation, normal T cell expressed and
secreted), phagocytosis ability or ROS (reactive oxygen species)
generation. Ionized serum calcium will also be determined on
samples collected on days 1, 7, 17, 18 and 21. Blood samples
collected on day 1 of arrival to the ARC and days 7, 8, 12, 17, 18
and 21 will be assayed for Heat Shock Protein (HSP) and cortisol.
These samples will be stored on ice until centrifugation
(15,000.times.g) for 15 minutes at 4.degree. C. Plasma will be
removed and stored at -20.degree. C. and the buffy coat fraction
will be removed and stored in Trizol at -80.degree. C. On days of
blood sampling, additional milk samples will be collected at both
the AM and PM milking and neutrophils isolated.
[0139] An additional possibility is the sampling of fecal cortisol
as a noninvasive measure of cortisol output. Another consideration
is to measure corticoid binding globuli (CBG). An increase in CBG
would reduce effective concentrations of cortisol and cause
increases in ACTH.
Example 4
[0140] Procedures:
[0141] Calf-fed steers (306 head; BW 263.5.+-.18.6 kg; 36 pens)
were utilized in a randomized block design experiment. Steers were
received over a two-day span at the feedlot. Upon arrival, steers
were allowed access to water and were processed, weighed, and
allocated to treatment within 12 h. During processing, steers were
identified with an individual ear tag, individually weighed,
vaccinated with Bovi-Shield Gold One Shot.TM. (IBR, BVD), Dectomax
Injectable.RTM. (internal and external parasiticide), and
Somubac.RTM. (Haemophilus somnus disease complex. Shrunk BW was a
single weight collected at time of processing following arrival.
Steers were blocked based on arrival date resulting in two blocks.
Within blocks, steers were assigned randomly to one of three
treatments by gate sorting every two steers to a pen; pen was then
randomly assigned to treatment. Treatments included: a control
group (Control): no supplementation with the disclosed composition
(OmniGen-AF.RTM. [OG] comprising between 15% and 40% silica,
between 50% and 81% mineral clay, between 1.0% and 5.0%
.beta.-glucans, between 0.05% and 3.0% .beta.-1,3
(4)-endoglucanohydrolase and between 1% and 8.0% mannan) OG
supplemented at 4 g/cwt of BW for 28 d (receiving period); and OG
supplemented at 4 g/cwt of BW for 215 d (the entire feeding
period). Supplementation of OG for both the 28 d and 215 d
treatment groups was formulated and added to the daily delivery of
the diet prior to feeding. A ninth steer was randomly added to nine
pens of the control group and the treatment group that only
received OG for 28 d during processing of the 2.sup.nd block. These
18 steers (9 hd/treatment group) would allow for the selection of
one steer from each of these 18 pens to be utilized for the
intravenous lipopolysaccharide (LPS) challenge portion of the
trial.
[0142] At the conclusion of the 28 d receiving period, steers were
limit fed for a period of 4 d at 2% BW to obtain an end of
receiving period BW. At the conclusion of the 28 d receiving
period, OG was no longer supplemented to the group of steers within
the treatment group that only received OG for the 28 d of the
receiving period; steers within the other OG treatment group (fed
for 215 d) continued to receive OG. OG supplementation for the
remaining treatment group was recalculated every 30 d to supply 4
g/cwt of BW on average. All steers were implanted with Revalor.RTM.
XS at the conclusion of the receiving period. During the last 28 d
of the finishing period, all cattle were supplemented
Optaflexx.RTM. ractopamine composition at a rate of 300 mg/hd/d. At
the end of the trial, steers were transported 51.5 km to a
commercial abattoir and held over-night. The following morning
steers were harvested; at which time hot carcass weights (HCW) were
recorded. Following a 48-h chill, fat thickness, LM area, and USDA
marbling score were determined. Final BW, ADG, and F:G were
calculated using HCW adjusted to a standard (63%) dressing
percentage.
[0143] To evaluate the immune response, two LPS challenges were
conducted; a subcutaneous LPS challenge (steers from the Control
group and the OG fed for 215 d group) and an intravenous LPS
challenge (steers from the Control group and the OG fed for 28 d
group). For the subcutaneous LPS, three pens from the Control and
the OG fed 215 d treatment group were randomly selected. From the
selected pens, 6 steers/pen were randomly selected and challenged
with LPS via subcutaneous injection on d 21. On d 21, steers from
selected pens were removed from the pen and moved to the processing
barn. As each steer was processed, BW was determined and an
indwelling rectal probe was inserted. After determining BW, each
steer was challenged with a subcutaneous injection of LPS at a rate
of 0.5 .mu.g/kg of BW and then returned to the feedlot pen. For the
intravenous LPS challenge, on d 25 of the receiving period, 18
steers (nine steers from Control and OG fed for 28 d treatment
groups) were randomly selected from the 9 Control and 9 OG fed for
28 d pens that contained 9 steers/pen. Steers were identified and
moved into a tie stall barn. After a 3 d adjustment period, steers
were fitted with indwelling jugular vein catheters for serial blood
collection and indwelling rectal temperature (RT) recording
devices, set to record RT at 1-min intervals continuously
throughout the immune challenge study. After insertion of the
jugular catheter and RT probe, steers were returned to the
individual tie stalls and allowed to rest for the remainder of the
d.
[0144] On the following day, from 0800 to 1600 h, blood samples
were collected at 30-min interval; two h prior to the challenge
(0800-1000 h) and six h after the challenge (1000-1600 h). At 1000
(0 h), following the collection of the blood sample, steers were
administered an i.v. dose of lipopolysaccharide (LPS, 0.5 .mu.g/kg
BW; purified from E. coli O111:B4; Sigma-Aldrich, St. Louis, Mo.).
A final blood sample was collected 24 h post LPS challenge. At each
collection point, 9 mL of blood was collected via monovette tubes
containing no additive for serum. After collection, blood samples
were allowed to clot for 30 min at room temperature and then
centrifuged at 2,000.times.g for 30 min (39.2.degree. F.). Serum
was collected and transferred into 1.5 mL microcentrifuge tubes and
stored at -112.degree. F. until analyzed. Serum was analyzed for
cortisol and pro-inflammatory cytokines (tumor necrosis
factor-.alpha., TNF-.alpha.; interferon-.gamma., IFN-.gamma.; and
interleukin-6, IL-6).
[0145] Feedlot performance data were analyzed as a randomized block
design using MIXED procedures of SAS (SAS Institute, Inc., Cary,
N.C.). Steers were blocked by arrival date and pen was the
experimental unit; model included the fixed effect of treatment and
block was a random effect. Immune response data were analyzed as a
completely randomized design with repeated measures using the MIXED
procedures of SAS; model included fixed effects of treatment and
time, treatment.times.time was used as the error term to test whole
plot effect. For both feedlot and immune data, when results of
F-test were significant (P<0.05), group means were compared by
use of least significant difference. Pair wise differences among
least squares means at various sample times were evaluated with the
PDIFF option of SAS. Distribution of USDA Quality Grade data were
analyzed as a randomized block design using the Glimmix procedure
of SAS.
[0146] Results:
[0147] For the receiving portion of the trial, there was no
difference in initial BW (P=0.82), ending BW (P=0.43), ADG
(P=0.32), DMI (P=0.76), and F:G (P=0.35) between the three
treatments (Table 2). In terms of morbidity related to respiratory
diseases, there was no difference (P=0.21) in the % of steers
treated (Table 3). There was also no difference in overall feedlot
performance; DMI (P=0.89), ADG (P=0.66) and F:G (P=0.90) were
similar across the three treatments (Table 2). There was no
difference in final BW (P=0.59), HCW (P=0.60), LM area (P=0.31),
marbling score (P=0.96), 12.sup.th rib fat thickness (P=0.86), or
calculated yield grade (P=0.52) between the three treatment groups.
The distribution of USDA Quality Grade was analyzed across all
three treatments. There was no difference in the amount of USDA
Prime, USDA Choice, or USDA Select Quality Grades for all three
treatments (Table 3). While there was no statistical difference in
terms of the percentage of carcasses grading USDA Choice or
greater, the economic significance is still of major importance.
Steers supplemented with OG for 215 d had a rate of USDA Choice
that was 81.91%, compared to 76.40 for Control steers. With a
$13.58 Choice/Select spread (11 Dec. 14), this difference in Choice
between the two treatment groups is the equivalent of .about.$850
difference in carcass value.
TABLE-US-00003 TABLE 3 Receiving period and overall feedlot
performance and carcass merit for steers fed no OG (CON), OG during
the receiving period (OG-28), or OG for 215 d (OG-215) Treatment
groups.sup.1 Item Con OG-28 OG-215 SEM P-value Receiving
Performance Initial BW (kg) 259 256 262 3.2 0.82 Ending BW
(kg).sup.2 300 304 306 3.6 0.43 DMI (kg/d) 7.5 7.7 7.6 0.4 0.76 ADG
(kg/d).sup.3 1.44 1.54 1.58 0.15 0.32 G:F.sup.4 0.9 0.9 0.09 --
0.35 Morbidity (%).sup.5 6.9 2.0 3.0 3 0.12 Feedlot Performance
Initial BW (kg) 261 572 572 6 0.87 Final BW (kg).sup.6 649 642 643
11 0.59 DMI (kg/d) 9.80 9.80 9.88 0.3 0.89 ADG (kg).sup.7 1.81 1.79
1.79 0.04 0.66 G:F 0.18 0.18 0.18 -- 0.90 Carcass Merit HCW (lbs)
409 405 405 3.2 0.96 LM area (in.sup.2) 94.78 91.36 94.06 1.61 0.31
Calculated YG 3.09 3.28 3.09 0.14 0.52 12.sup.th rib fat (cm) 1.37
1.42 1.40 0.08 0.86 Marbling.sup.8 503 498 508 24 0.96 Prime (%)
4.49 2.13 5.32 2.32 0.53 Choice (%) 76.40 76.60 81.91 4.50 0.59
Select (%) 19.10 21.28 13.83 4.22 0.41 .sup.1CON: No OG; OG-28: OG
during receiving, OG: OG for 215 d. .sup.2Limit fed at 2% of BW for
4 d prior to single BW to determine ending BW of receiving period
.sup.3Calculated from ending BW of receiving period .sup.4Analyzed
as G:F, the reciprocal of F:G. .sup.5Overall percentage of steers
treated for bovine respiratory disease .sup.6Calculated from
carcass weight, adjusted to 63% common dressing percent. .sup.7ADG
for the entire feeding period (including receiving period)
.sup.8Marbling Score: 400 = Small, 500 = Modest, etc.
[0148] For the subcutaneous LPS challenge, there was a treatment
(P=<0.001) and time (P=<0.001) effect for RT (FIG. 13). Both
treatment groups responded to the subcutaneous LPS challenge, with
both treatment groups achieving a maximum RT approximately 5 h post
challenge (Control=40.10.degree. C. vs. OG=39.92.degree. C.; FIG.
13). Furthermore, both maintained similar patterns (P=1.00)
throughout the subcutaneous LPS challenge. Steers in the Control
treatment group had an average RT that was greater than the OG
treatment group (39.22.degree. C. and 39.07.degree. C.,
respectively). There was no treatment effect (P=0.75) or
treatment.times.time interaction (P=0.99) for change in RT from
baseline (FIG. 14). Again, there was a treatment effect for RT
(P=<0.001), but this treatment difference was not observed when
comparing the change in RT from baseline between the treatment
groups (average change from baseline was 0.03.degree. C. for
Control and 0.03.degree. C. for OG).
[0149] For the intravenous LPS challenge portion of the trial,
there was a treatment (P=0.002) and time effect (P=<0.001),
however there was no treatment.times.time interaction (P=0.99) for
RT (FIG. 15). Steers supplemented with OG had a greater RT when
compared to Control (39.26.degree. C. vs. 39.17.degree. C.,
respectively) during the intravenous challenge. This average RT is
very similar to the RT that was observed for the steers that were
utilized in the subcutaneous LPS challenge. In terms of response to
the intravenous LPS challenge, both treatment groups responded to
the LPS challenge similarly (P=0.99). For both groups of steers,
maximum RT was observed 2.5 h post intravenous LPS administration,
and within 6 h, RT had returned to baseline temperatures (-21 to 0
h prior to the challenge: FIG. 15). There was also no treatment
effect (P=0.49) or treatment.times.time interaction (P=0.99) for
change in RT from baseline (-21 to 0 h: FIG. 16). Baseline line
temperatures were different (P<0.001) between the two treatment
groups, but the change from baseline was similar (P=0.49) between
the Control and OG supplemented steers (FIG. 16).
[0150] For serum concentrations of cortisol, there was a treatment
(P=0.005) and time effect (P=<0.001); though there was no
treatment.times.time interaction (P=0.99). Steers supplemented OG
had decreased concentrations of cortisol, when compared to the CON
steers (25.5 vs 29.2 ng/ml, respectively; FIG. 17). In terms of
response to the intravenous LPS challenge, both treatment groups
responded similarly (P=0.99). Prior to the LPS challenge, both
treatment groups had cortisol concentrations below 10 .mu.g/mL.
Thirty min after the LPS challenge, cortisol concentrations had
increased (P=<0.001) to above 35 .mu.g/mL; cortisol
concentrations for both treatment groups did not return to baseline
concentrations until 24 h post LPS challenge (FIG. 18).
[0151] Blood urea nitrogen (BUN), non-esterified fatty acids, and
glucose were analyzed to evaluate metabolic alterations during the
intravenous LPS challenge. For BUN's, there was a treatment
(P=<0.001) and time (P=<0.001) effect, but there was no
treatment.times.time interaction (P=0.99). Control steers had a
greater (P=<0.001) concentration of BUNs when compared to the OG
supplemented steers (FIG. 19). This difference in BUNs was observed
prior (P=0.01) and post LPS challenge (P=<0001). Regardless of
the LPS challenge, Control steers' BUN concentrations were on
average 12.4 mg/mL while OG-supplemented steers' BUN concentrations
were on average 11.5 mg/mL (FIG. 19.) There was a treatment
(P=0.009) and time (P=<0.001) effect for glucose concentrations;
no treatment.times.time interaction (P=0.19). Serum glucose
concentrations were greater (P=<0.001) for OG-supplemented
steers, when compared to Control steers (76.4 mg/mL vs. 72.4 mg/mL,
respectively; FIG. 20). This difference in glucose was not apparent
prior to the LPS challenge, as baseline concentrations (-2 to 0 h)
were similar (0.62; FIG. 21). However, following the LPS challenge,
while both treatment groups had a decrease in serum glucose, this
reduction in glucose was greater (P=0.009) in the Control steers,
when compared to the OG-supplemented steers (FIG. 21). Prior to the
LPS challenge, both treatment groups maintained serum glucose
concentrations greater than 75 mg/mL. However, following the LPS
challenge (1 h post LPS challenge) serum glucose concentrations
started to decrease, and continued to decrease until 3.5 h after
administrations of the LPS challenge (FIG. 21). During this
decrease in glucose concentrations, concentrations in the Control
steers decreased to a greater extent than the OG-supplemented
steers. While this decrease of glucose concentrations did not
result in a treatment.times.time interaction, this decrease did
result in an overall treatment difference between the two treatment
groups (FIG. 21). There was a treatment (P=<0.001) and time
(P=<0.001) effect, and a tendency for a treatment.times.time
interaction (P=0.007) for serum NEFA concentrations. Steers within
the Control treatment group had greater (P=<0.001) serum NEFA
concentrations, when compared to OG-supplemented steers (FIG. 22).
This difference in NEFA concentrations was present prior (P=0.006)
to the intravenous LPS challenge. However, following the LPS
challenge these differences were even greater (P=<0.001: FIG.
22). Prior to the LPS challenge, NEFA concentrations for Control
steers were 0.10 mmol/ml while OG-supplemented steers were 0.07.
However, following the LPS challenge, NEFA concentrations in the
Control steers increased to 0.23 mmol/mL while the OG-supplemented
steers only increased to 0.11 mmol/mL.
[0152] There was a treatment (P=<0.001) and a time effect
(P=<0.001) for the pro-inflammatory cytokine IFN-.gamma.; there
was no treatment.times.time interaction (P=0.77). OG-supplemented
steers had a greater production of IFN-.gamma., when compared to
the Control steers (FIG. 23). Prior (P=0.05) and post (P=<0.001)
LPS challenge, the concentrations of IFN-.gamma. were greater in
the OG-supplemented steers, when compared to the Control steers
(FIG. 23). There was also a treatment (P=0.03) and time
(P=<0.001) effect for TNF-.alpha.; there was no
treatment.times.time interaction (P=0.42). Overall serum
concentrations for TNF-.alpha. were greater (P=0.03) in the
OG-supplemented steers, when compared to the Control steers (FIG.
24). For both treatment groups, prior to the LPS challenge, no
detectable concentrations of TNF-.alpha. were present, and this no
detectable concentrations continued until 0.5 h post challenge
(FIG. 25). At 0.5 h, concentrations of TNF-.alpha. dramatically
increased (P=<0.001) for both treatment groups. Concentrations
of TNF-.alpha. returned to near baseline (-2 to 0 h) around 4 h
after the LPS challenge (FIG. 25). There was only a time effect
(P=<0.001) for the pro-inflammatory cytokine IL-6 (FIG. 26). As
with TNF-.alpha., prior to the LPS challenge, concentrations of
IL-6 were not detectable. However, 1 h after the LPS challenge,
concentrations of IL-6 dramatically increased (P=<0.001);
reaching maximum concentrations at 2 h post LPS challenge. From 1
to 8 h post LPS challenge, IL-6 concentrations remained elevated
above baseline concentrations, and did not return to baseline
concentrations until 24 h post LPS challenge (FIG. 26).
[0153] The mean complete blood cell count (CBC) analysis is
reported in Table 4. Supplementation of OG did not impact red blood
cell counts (P=0.45) or the monocyte percentage (P=0.26).
Hemoglobin concentrations (P=0.008), Hematocrit % (P=0.03), and
white blood cell concentration (P=0.001) were greater (P=0.008) in
Control steers, when compared to OG-supplemented steers. Neutrophil
% (P=0.04) and eosinophils concentration (P=0.02) were greater in
OG-supplemented steers, when compared to Control steers. For the %
of lymphocytes, there was a treatment.times.time interaction
(P=0.006). Prior to the LPS challenge (-2 to 0 h), Control steers
had a greater (P=<0.001) concentration of lymphocytes compared
to the OG-supplemented steers (FIG. 27). One h after the challenge,
there was no difference (P=0.35) between the treatment groups, and
from 1 h to 8 h, there was no difference (P=0.66). However, 24 h
after the LPS challenge, Control steers had a greater (P=0.005)
concentration of lymphocytes (FIG. 27). There was also a
treatment.times.time interaction (P=0.006) for the
neutrophil:lymphocyte ratio. Prior to the LPS challenge, the
neutrophil:lymphocyte ration was greater (P=<0.001) for OmniGen
steers, when compared to Control steers (FIG. 28).
TABLE-US-00004 TABLE 4 Mean Complete Blood Cell Count values newly
received steers supplemented OG at a rate of 4 mg/cwt during the
receiving period (OMN-28) or no OmniGen - AF (CON) during a
lipopolysaccharide challenge P-value Variables OmniGen Control SEM
TRT TIME TxT Red Blood Cells (mil/.mu.L) 8.46 8.47 0.30 0.45 0.41
0.99 Hemoglobin (g/dL) 10.61 10.97 0.09 0.008 0.18 0.99 Hematocrit
(%) 31.06 32.29 0.38 0.03 0.46 0.99 White Blood Cells (K/.mu.L)
4.47 5.42 0.12 0.001 <0.001 0.88 Neutrophils (%) 0.43 0.26 0.05
0.04 <0.001 0.34 Lymphocytes (%) 3.02 3.63 0.09 <0.001
<0.001 0.006 Monocytes (%) 1.06 1.14 0.20 0.26 <0.001 0.23
Eosinophils (K/.mu.L) 0.07 0.04 0.005 0.02 0.76 0.76
Neutrophils:Lymphocytes 0.14 0.09 0.016 0.01 <0.001 0.006
[0154] Conclusions:
[0155] In this study, newly received calf-fed steers supplemented
with OG for a period of 28 d (receiving period) or for 215 d
(entire feeding period) did not impact feedlot performance or
carcass merit. While there was no statistical difference in terms
of the percentage of carcasses grading USDA Choice or greater, the
economic significance should outweigh the lack of a statistical
difference. With a $13.58 Choice/Select spread (11 Dec. 2014), this
difference in Choice between the two treatment groups is the
equivalent of .about.$850 in carcass value. Furthermore,
supplementation of OG alters the immune response. When challenged
with a lipopolysaccharide, OG supplementation appears to prime the
pro-inflammatory response (as evident by increased concentrations
of IFN-.gamma. and TNF-.alpha.). In addition, there was the
difference in metabolism that was observed. The increased
concentrations of BUN's and NEFA's within the Control steers
indicate a possible greater energy need to initiate and sustain an
immune response, when compared to the OG-supplemented steers. This
increase in greater energy demand is further indicated as the
OG-supplemented steers had greater concentrations of serum glucose,
when compared to the Control steers. Overall, these results suggest
that supplementation of OG may enhance the immune response of
calf-fed steers upon feedlot entry.
Example 5
[0156] A study was conducted to identify genes expressed by
circulating immune cells that are regulated by a commercial
embodiment of Composition I. Rats (n=6 per group) were randomly
assigned to Composition I and control groups. Composition I was
supplemented in the diet at 0.5% in the Composition I group. Total
RNA was purified from whole blood and gene expression was analyzed
with the use of the Rat Innate and Adaptive Immune Responses RT2
Profiler Polymerase Chain Reaction (PCR) Array (SABiosciences,
Qiagen). A total of 84 target genes were present on the array. Gene
expression of circulating immune cells was analyzed at seven,
fourteen, twenty-one and twenty-eight days of Composition I
supplementation. The expression of 67 genes changed following
Composition I supplementation across the time points. Table 5 lists
the genes with altered gene expression following Composition I
supplementation and includes information indicating stimulation (+)
or repression (-) of gene expression.
TABLE-US-00005 TABLE 5 Gene Repressed Gene Induced Reference Genes
Crp - Ifnb1 + Actb Mbl2 - Cd80 + Ldha Apcs - Tlr1 + Rplp1 Il5 -
Tlr6 + Ifna1 - Mapk8 + Ccl12 - Nod2 + Csf2 - Ccr8 + Il13 - Irak1 +
Il10 - Cd1d1 + Gata3 - Stat4 + Stat3 - Il1r1 + C3 - Faslg + Tlr3 -
Irf3 + Ccl5 - Ifnar1 + Mx2 - Slc11a1 + Nfkb1 - Tlr4 + Nfkbia - Cd86
+ Tlr9 - Casp1 + Cxcl10 - Ccr5 + Cd4 - Icam1 + Il6 - Camp + Ccl3 -
Tlr7 + Ccr6 - Irf7 + Cd40 - Rorc + Ddx58 - Cd40lg + Il18 - Tbx21 +
Jun - Casp8 + Tnf - Il23a + Traf6 - Cd14 + Stat1 - Cd8a + Cxcr3 +
Foxp3 + Lbp + Mapk1 + Myd88 + Stat6 + Agrin + IL33 +
Example 6
[0157] Feeding OmniGen-AF.RTM. (Composition I or OG; Prince Agri
Products, Inc., Quincy, Ill.) at 0.5% of the diet supports immune
function in ruminant livestock. Targeted profiling of
immune-associated genes in whole blood is an established
methodology to evaluate the efficacy of feed additives with
immune-altering properties. Higher daily inclusion rate of OG than
0.5% may be required to optimize immune function. The objective of
this study was to evaluate the effect of dietary OG inclusion rate
(1% vs. 0.5%) on the expression profile of immune-associated genes.
Male CD rats (5/treatment) weighing 180-200 grams had ad libitum
access to a diet with 0 (control), 0.5 (1.times.), or 1% (2.times.)
of OmniGen-AF.RTM. for 28 days. At the end of the feeding period,
whole blood was collected. RNA was purified from whole blood
samples and used to generate cDNA that acted as template in the Rat
Innate and Adaptive Immune Responses RT.sup.2 Profiler PCR array
(SABiosciences). Using PROC GLM, cDNA abundance of
immune-associated genes was compared between control and
supplemented groups (0.5 or 1%) with a P<0.05 cut-off value for
significance. Of the 79 immune-associated genes that were expressed
above the detection limit in all samples, 16 (7 up-regulated) and
13 genes (8 up-regulated) were altered by 0.5% and 1% OG
supplementation, most of which (11 with 6 up-regulated) were
altered at both OG inclusion rates. Genes that were up-regulated at
both rates include IL13 (0.5%: +3.16, 1%: +3.70 fold-change), IL5
(0.5%: +2.64, 1%: +2.62), Irak1 (0.5%: +2.50, 1%: +1.98), Nod2
(0.5%: +1.83, 1%: +2.02), IFNa1 (0.5%: +1.81, 1%: +2.10), and Cd80
(0.5%: +1.77, 1%: +2.47). Genes that were down-regulated at both
inclusion rates include TLR3 (0.5%: -2.22, 1%: -2.39), CxCL10
(0.5%: -2.19, 1%: -2.26), STAT1 (0.5%: -2.07, 1%: -1.99), STAT3
(0.5%: -2.05, 1%: -1.92), and NF.kappa.b1 (0.5%: -1.84, 1%: -1.75).
The results suggest that OG supplementation inclusion rate
independently promotes immune function through various pathways
including pathogen recognition, adaptive immune cell activation,
and various transcription factors.
Example 7
[0158] The OmniGen-AF.RTM. composition (Composition I or OG; Prince
Agri Products, Inc., Quincy, Ill.) is a branded proprietary product
shown to augment immune function in ruminants and other species.
Targeted profiling of immune-associated genes in whole blood is an
effective platform for identification of multiple immune response
markers to feed additives with immune-altering properties. The
objective of this study was to identify multiple immune response
markers that are increased by dietary OG throughout a 28-d
supplementation period. It was hypothesized that several
immune-associated genes in whole blood are consistently
up-regulated during a 28-d supplementation period. Fourteen male CD
rats weighing 180-200 grams had ad libitum access to a diet
containing 0 (control; n=5, only 28 days) or 0.5% OG for 7 (n=4) or
28 days (n=5). Whole blood was collected at the end of the feeding
period. RNA was purified from whole blood samples and used to
generate cDNA that acted as template in the Rat Innate and Adaptive
Immune Responses RT.sup.2 Profiler PCR array (SABiosciences). Using
PROC GLM, we compared cDNA abundance of immune-associated genes
between control and supplemented groups (7 or 28 d) with a
P<0.05 cut-off value for significance. Of the 77
immune-associated genes that were expressed above the detection
limit in all samples, 6 genes were up-regulated after 7 d of OG
supplementation and 4 genes were up-regulated after 28 d of OG
supplementation. Three genes were up-regulated after 7 d (Cd80:
+2.40; Irak1: +2.25; Nod2: +2.08 fold-change) as well as after 28 d
of OG supplementation (Cd80: +1.77; Irak1: +2.50; Nod2: +1.83
fold-change). The results suggest Cd80, Irak1, and Nod2 as immune
response markers that are increased by dietary OG throughout a 28-d
supplementation period.
Example 8
[0159] This study was designed to determine the effect of
supplementing feedlot steers with the OmniGen-AF.RTM. composition
(Composition I, OG) on the acute phase response to a
lipopolysaccharide (LPS) challenge. Steers (n=18; 270.+-.5 kg BW)
were separated into two treatment groups (n=9/treatment): one group
was fed a standard receiving diet (Control, Cont) and the other
group was fed the same receiving diet supplemented with OG at 4
g/45.4 kg BW for 29 d (OG). On d27 steers were fitted with
indwelling jugular cannulas and rectal temperature (RT) monitoring
devices and placed in individual stalls. On d28, steers were
challenged i.v. with LPS (0.5 .mu.g/kg BW at 0 h). Sickness
behavior scores (SBS) and two whole blood samples were collected at
30-min intervals from -2 to 8 h relative to the challenge at 0 h.
One vacutainer containing EDTA was collected for complete blood
cell count (CBC) analysis, and the second was collected in 9-mL
monovette serum tube; after collection serum was isolated and
stored at -80.degree. C. until analyzed for cortisol and cytokine
concentrations. Rectal temperature, SBS, and cortisol were affected
by time (P<0.001). Prior to the challenge, RT was greater
(P<0.001) in Cont steers (39.31.+-.0.03.degree. C.) than OG
steers (39.14.+-.0.03.degree. C.). Therefore, post-challenge RT was
analyzed as the change in response from baseline values. The change
in RT relative to baseline values increased (P<0.001) in both
groups in response to LPS challenge, but was not affected by
treatment (P=0.49). Sickness behavior scores increased (P<0.001)
after LPS challenge and tended (P=0.09) to be greater in Control
(1.57.+-.0.02) than OG steers (1.51.+-.0.02). Cortisol
concentrations were affected by treatment (P=0.005) and time
(P<0.001). For both groups, cortisol increased (P<0.001) in
response to LPS challenge. Cortisol was greater in Cont
(25.2.+-.0.9 ng/mL) than OG steers (25.5.+-.0.9 ng/mL). White blood
cell and lymphocyte concentrations were greater (P<0.004) in
Cont than OG steers throughout the study. Neutrophils were
decreased (P=0.04) in Cont steers (0.7.+-.0.2 K/uL) compared to OG
steers (1.3.+-.0.2 K/uL) prior to the LPS challenge. There was a
treatment (P<0.02) and time (P<0.001) effect for tumor
necrosis factor-.alpha. (TNF.alpha.) and interferon-.gamma.
(IFN.gamma.). Specifically, TNF.alpha. and IFN.gamma.
concentrations increased (P<0.001) in response to LPS challenge.
Furthermore, concentrations of TNF.alpha. and IFN.gamma. were
decreased in (P<0.02) in Cont steers compared to OG steers.
These data suggest that OG supplementation served to prime the
immune system prior to the LPS challenge, allowing for an enhanced
response to LPS challenge.
Example 9
[0160] The use of probiotic feed supplements to enhance animal
health and growth are of great interest to the beef industry.
Studies have demonstrated that some probiotic supplements may
affect metabolism, and therefore influence an animal's response to
an immune challenge. This study was designed to determine the
effect of supplementing feedlot steers with OmniGen-AF.RTM.
composition (Composition I, OG) during the receiving period on the
metabolic response to a lipopolysaccharide (LPS) challenge. Steers
(n=18; 270.+-.5 kg BW) were obtained and transported to the
feedlot. Upon arrival steers were processed and separated into 2
treatment groups (n=9/treatment): one group was fed a standard
receiving diet (Control; Cont) and the other group was fed the same
receiving diet supplemented with the OmniGen-AF.RTM. composition at
4 g/45.4 kg BW/d for 29 d (OG). On d 27 steers were fitted with
indwelling jugular cannulas and placed in individual stalls. On d
28, steers were challenged i.v. with LPS (0.5 .mu.g/kg BW at 0 h)
and blood samples were collected at 30-min intervals from -2 to 8 h
and at 24 h post-challenge. Serum was isolated and stored at
-80.degree. C. until analyzed for glucose, non-esterified fatty
acids (NEFA) and blood urea nitrogen (BUN) concentrations. Glucose
concentrations were affected by treatment (P=0.009) and time
(P<0.001). Glucose was greater in OG steers compared to Cont
steers (76.4.+-.1.1 mg/dL vs. 72.4.+-.1.0 mg/dL). For NEFA
concentrations, there was a treatment (P<0.001) and time
(P<0.001) effect. Specifically, Cont (0.210.+-.0.007 mmol/L)
steers had greater NEFA concentrations than OG steers
(0.101.+-.0.010 mmol/L). There was a tendency (P=0.07) for a
treatment.times.time interaction such that NEFA concentrations were
greater (P<0.03) in Cont steers than OG steers from 3 to 8 hr
after LPS challenge. For BUN, there was a treatment (P<0.001)
effect such that concentrations were greater in Cont steers
(12.4.+-.0.1 mg/dL) than OG supplemented steers (11.5.+-.0.1 mg/dL)
throughout the study, and were not affected by time (P=0.28). These
data suggest that OG supplementation modulates the metabolic
response to a LPS challenge and provides an indication that
supplementation of feedlot steers with OG may prevent the breakdown
of other substrates (e.g., protein and fat) for energy during an
immune challenge.
[0161] In view of the many possible embodiments to which the
principles of the disclosed technology may be applied, it should be
recognized that the illustrated embodiments are only preferred
examples of the technology and should not be taken as limiting the
scope of the technology. Rather, the scope of the technology is
defined by the following claims. We therefore claim as our
invention all that comes within the scope and spirit of these
claims.
* * * * *