Note: Descriptions are shown in the official language in which they were submitted.
CA 02470417 2004-06-09
USE OF CORTICOSTEROIDS ALONE OR IN CONJUNCTION WITH
NUTRITIONAL THERAPY TO REDUCE STRESS IN ANIMALS
FIELD OF THE INVENTION
The invention pertains to the use of corticosteroids alone, or in conjunction
with a
nutritional supplement to alleviate stress in animals.
BACKGROUND OF THE INVENTION
Transport and handling to which animals are exposed during standard rearing
and
l0 marketing practices can be stressful. A plethora of effects has been
reported including
hypoglycemia, dehydration, energy depletion, protein degradation, electrolyte
imbalance and
stimulation of the hypothalamic-pituitary-adrenal (HPA) axis (Schaefer et al.,
1997; 2001).
Such practices may contribute to significant weight loss in addition to other
negative effects
in animals.
~5
The attenuation of the effects of transport and handling stress has typically
involved
the examination of the stressors themselves; for example, improvements in
animal loading
facilities, transport earners, shock absorbers, wind protection, and loading
densities have
been recommended, but met with varying levels of success.
An alternative approach involves treatment of the animal rather than the
stressor.
Attempts have been made to treat the effects of transport stress with a wide
variety of
different compounds, including amino acids (Japanese Patent Application No. 61-
280239 to
Takaaki et al.); epinephrine (United States Patent No. 6,133,321 to Prusa et
al.); ethyl alcohol
(Japanese Patent Application No. 01-113669 to Kosaku et ad.); glycerin,
licorice roots,
vitamins C and E (Russian Patent Nos. 2,160,532, 2,160,533 and 2,153,802 to
Ehzergajl et
al.); human interleukin-2 (United States Patent No. 4,818,769 to Nunberg, et
al.); ionol
(Russian Patent No. 2,147,799 to Ajtuev et al.); sedatives such as
acepromazine, xylazine and
pentobarbitone (Brearly et al.,1990; Sanhouri et al,, 1991 ); and sorbitol
(United States Patent
No. 5,137,735 to Bignon). In addition, different nutritional therapies have
been reported to be
effective in reducing transport and handling stress (Schaefer et al., 1997;
2001 ). United
States Patent Nos. 5,505,968 and 5,728,675 to Schaefer et al. issued April 9,
1996 arid March
CA 02470417 2004-06-09
17, 1998 respectively, relates to a nutritional supplement for animals to
prevent or reduce
antemortem staress.
Reducing weight loss and other deleterious effects in animals during transport
or
handling stress contributes to both animal welfare and profitability. As
existing practices and
the prior art do not address the issue of weight loss in animals during
transport, there is
evidently a need for an effective method or therapy that achieves alleviation
of transport
stress in animals. Further, commercially available corticosteroids are
typically used to treat
inflammatory diseases or other illnesses in animals, but current practices for
alleviating stress
1o in animals do not involve the use of cortiosteroids alone or in conjunction
with a nutritional
supplement.
SUMMARY OF THE INVENTION
The inventors discovered that provision of a corticostc:roid to an animal
mitigates the
effects of stress, particularly weight loss, when a therapeutically effective
amount of the
corticosteroid is administered to the animal in need thereof. The inventors
found that the
administration of the corticosteraid in conjunction with a nutritional
supplement to the animal
provides synergistic effects in alleviating the effects of stress. The
combined administration
of a carticosteroid and nutritional supplement also was found to have a
restorative effect
when administered following transport.
Broadly stated, the invention provides a method of allE:viating the effects of
stress in a
non-human animal comprising administering a therapeutically effective amount
of a
corticosteroid to the animal in need thereof. The invention extends to a
method of further
administering a corticosteroid in conjunction with a nutrient supplement to
the animal. The
nutrient supplement comprises:
a) one or more sources of electrolytes providing each of sodium, potassium and
magnesium;
b) one or more sources of amino acids providing each of alanine, lysine,
3o phenylalanine, methionine, threonine, leucine, isoleucine, valime, and
glutamate; and
c) a source of tryptophan.
2
CA 02470417 2004-06-09
As used herein and in the claims, the terms and phrases set out below have the
meanings which follow:
"Animal" refers to a non-human animal including, but not limited to, domestic
ruminant and monogastric animals, including swine (Sus dorraesticus), horses,
cattle (Bos
taccrus and Bos iredicus); domestic ungulates, including bison, sheep, Iamb,
deer, moose, elk,
caribou and goats; domesticated fowl, including chickens, turkeys, geese,
ducks, game birds,
and other birds raised in domestication to produce eggs or meat; dogs, cats
and other
companion animals; wild animals; captured animals; and zoo animals.
"Animal in need thereof ' means an animal prior to exposure to the stress,
during the
stress, and/or following exposure to the stress.
"Antemortem stress" means the stresses imparted to animals during pre-
slaughter
treatment, including transport, holding, management, and handling.
"Bypass form" means amino acids provided from feed sources such as bypass,
chelated or protected proteins. In such forms, the amino acids are not
substantially degraded
in the rumen of ruminant animals, but pass through to the abomasum
comparatively intact. In
2o general, in the context of this invention, an amino acid is considered to
be in a bypass form if
greater that about 40% of the protein in that feed source is in a bypass form.
Without
limitation, preferred feed sources of the bypass amino acids include
distillers grain, alfalfa
meal, corn gluten meal, skim milk powder, whey powder, soybean, caesin,
cottonseed meal,
feather meal, blood meal, bone and meat meal, and fishmeal.
"Carrier" means a suitable vehicle that is biocompatible and pharmaceutically
acceptable, including for instance, one or more solid, semisolid or liquid
diluents, excipients,
adjuvants, flavours, or encapsulating substances which are suitable for
administration.
"Corticosteroids" or "corticosteroid" means the C21 steroid hormones produced
by
the adrenal cortex. The term also refers to natural analogs and synthetic
equivalents of
corticosteroids including most preferably, dexamethasone, prednisone,
prednisolone, 6oc-
3
CA 02470417 2004-06-09
methylprednisolone, fludrocordsone, triarncinolone, paramethasone,
betamethasone,
aldosterone, and pharmaceutically acceptable salts thereof.
"Feed" means products including, but riot limited to, grasses, legumes,
grains, oil
seeds, forties, and sedges, for example oats, barley, wheat, canola, rye,
sorghum, millet, corn,
molasses, alfalfa, clover, brome, timothy or fescue, Bermuda grass, orchard
grass, rice straw,
and other suitable feedstuffs. The term also includes the above feed sources
of the bypass
amino acids.
"Hypotonic" means concentration of an ingredient, primarily related to the
concentrations of the electrolytes, in an amount that is not significantly
greater than the
concentration of that ingredient found in the physiological fluids of the
animal such as
plasma, interstitial and intracellular fluids (i.e. the isotonic
concentration). This concentration
is preferred so that the supplement provided to the animal will have a lower
osmotic pressure
in respect of the salts than that of the physiological fluids. Since many
animals experiencing
stress are dehydrated, the nutrient supplement is preferably formulated to
avoid hypertonic
liquids (or solids which will result in hypertonic concentrations). Hypertonic
solutions would
simply draw more fluid from the tissue and exacerbate tissue loss.
"Non-steady state" means a condition in which an animal's endocrine,
physiological
or metabolic values are in a state of flux often due to environmental factors
such as stressors.
"Pharmaceutically- or therapeutically-acceptable" means a substance that does
not
significantly interfere with the effectiveness of the corticosteroid, or
corticosteroid in
conjunction with a nutritional supplement and which has an acceptable toxic
profile for the
animal to which it is administered.
"Pharmaceutically-acceptable salts" means derivatives of the free acid or base
forms
of the corticosteroids that are modified by addition of appropriate salts.
"Steady-state" means a condition in which an animal's endocrine, physiological
and
metabolic values are all within a normal range and the animal i.s not
stressed.
4
CA 02470417 2004-06-09
"Stress" means environmental stresses imparted to animals during transport
stress,
antemortem stress, regrouping stress, handling and management stress, mixing
of animals,
post-parturient stress, weaning stress, acute weather stressors, noise
stressors, and when the
animal is in a non-steady state. The term also includes other similar animal
management
stresses.
"Therapeutically effective amount" means any amount of a formulation of the
corticosteroid alone, or corticosteroid in conjunction with a nutritional
supplement that is
sufficient to alleviate effects of stress when administered to the animal in
need thereof.
"Transport stress" means stresses imparted to animals. during transportation,
including
for example, handling, weighing, mixing and prods prior to loading; and noise,
air
temperature, velocity, crowding, pollutants, infectious agents., vibration,
motion, and
withdrawal of food and water during transport.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
The invention broadly provides a method of alleviating the effects of stress
in a non-
human animal comprising administering a therapeutically effective amount of a
corticosteroid
to the animal in need thereof. Most preferably, dexamethasone is used to
alleviate the effects
of stress, particularly weight loss. Weight loss generally occurs, for
example, during
transportation which exposes an animal to a variety of stressors, including
handling,
weighing, mixing and prods prior to loading; and noise, air temperature,
velocity, crowding,
pollutants, infectious agents, vibration, motion, and withdrawal of food and
water during
transport. Dehydration and tissue degradation contributes to significant
weight loss in an
animal during prolonged transport.
When an animal is exposed to such stressors, it typically responds by
displaying
activation of the hypothalamic-pituitary-adrenal (HPA) axis, resulting in a
substantial
increase in the production of an endogenous corticosteroid, namely cortisol
(Schaefer and
3o Cook, 2003). Corticosteroids (e.g., cortisol) are steroid hormones produced
by the adrenal
cortex, and function in metabolic events which are essential for life,
including carbohydrate,
protein and lipid metabolism; fluid and electrolyte dynamics; energy
procurement; and the
5
CA 02470417 2004-06-09
functions of the cardiovascular system, kidney, skeletal muscle, nervous
system and other
organs and tissues (Merck, 1973; Haynes, 1990). Cortisol influences numerous
biochemical,
endocrine and physiological functions to maintain homeosta sis of the animal.
However, this
response subsequently declines under negative feedback control, as cortisol
levels are often
reported to be relatively low following long-distance transport, indicating
fatigue of the HPA
axis.
However, the inventors have found that when an animal is supplemented with a
corticosteroid having greater potency and biological half-life than cortisol,
it is better able to
cope physiologically and exhibits less weight loss incurred by stress.
Corticosteroid
administration in accordance with this invention is most effective for
situations in which the
animal is exposed to high-energy dependent stressors for prolonged periods
(i.e., hours to
days). Exposure to cold or hot temperatures and physical activity (i.c.,
maintaining balance),
for example, typically occur during transport. Throughout this period, the
ability of the
animal's body to supply energy substrates or glucose is reduced due to
insufficient levels of
cortisol. Without being bound by the same, it is believed that administration
of a
corticosteroid supplements endogenous corticosteroid levels, and maintains
higher levels of
energy substrates during periods of peak energy demand.
Dexamethasone is regarded has being approximately 20 times as potent as
cortisol in
glucocorticoid activity and has a much longer biological half-life (Cook et
al., 1993).
Dexamethasone is commonly incorporated into pharmaceutical compositions as an
anti-
inflammatory or anti-infection agent (Haynes, 1990); anti-viral treatment
(Chinese Patent No.
1245058 to Zeng et al.); inducer ofparturition and lactation (United States
Patent No.
3,856,955 to Anderson; United States Patent No. 3,966,927 to Binninger);
growth promoter
(Meadus et al., 2002, J. Anim. Sci. 74:81-87; United Kingdom Patent No.
2,198,351 to
Wilson et al.); alleviator of acute surgical stress (Goodin et al., United
States Patent No.
5,670,495); and anti-nausea agent (Tyers et al., United States Patent No.
6,544,550).
However, to the inventors' knowledge, the prior art does not report use of a
corticosteroid, such as dexamethasone alone, or in conjunction with a
nutritional supplement,
to reduce transport stress, particularly the subsequent reduction in weight
loss. In the present
6
CA 02470417 2004-06-09
invention, dexamethasone was proven effective when administered to calves
prior to transport
to provide energy substrates for a longer period during transport than
achieved by the
endogenous response. The endogenous response was suppressed by the negative
feedback
action of dexamethasone on the HPA axis, but the glucocorticoid effects of
endogenous
cortisol were replaced by the much greater effects of the more powerful,
longer acting
dexamethasone.
Surprisingly, the inventors found that optimal synergistic effects are
obtained when
the corticosteroid is used in conjunction with a nutritional supplement. The
corticosteroid
to assists in supplying energy substrates, as mediated by catabolic breakdown
of proteins and
triglycerides, and Na+ ion retention. However, a nutritional supplement
contributes essential
amino acids and electrolytes necessary to augment the action of the
corticosteroid. Further,
supplementation with a nutritional supplement appears to facilitate an
animal's recovery
following transport, in that the animal displays one or more behavioral
changes, fvr example,
15 an improvement in appetite, and a decline .in lying frequency.
Corticosteroids, for the purposes of this invention, includes the C21 steroids
of the
adrenal cortex (for example, cortisol, corticosterone, and aldosterone),
natural analogs, and
most preferably, synthetic equivalents of corticosteroids including
dexamethasone,
2o prednisone, prednisolone, hoc-methylprednisolone, fludrocortisone,
triamcinolone,
paramethasone, betamethasone, aldosterone, and pharmaceutically-acceptable
salts thereof.
Corticosteroids are broadly classified as mineralocorticoid or glucocorticoid
in accordance
with their major function. MineraIocorticoids control salt and water balance
through action on
the kidneys, whereas glucocorticoids influence synthesis, storage and
utilization of glucose.
25 Most corticosteroids display one type of activity or have varying degrees
of both types of
activity; for example, aldosterone and desoxycorticosterone are highly potent
mineralocorticoids, but lack glucocorticoid activity. Cortisol and cortisone
are naturally
occurring glucocorticoids, yet have weak mineralocorticoid activity.
Corticosterone has
modest activities in both categories (Haynes, 1990). It1 the prior art,
corticosteroids are
30 primarily used as anti-inflammatory and immunosuppressive agents to treat
both humans and
animals (Haynes, 1990; Merck, 1973). The relative potency of corticosteroids
in terms of
7
CA 02470417 2004-06-09
their mineralocorticoid, glucocorticoid and anti-inflammatory actions can be
expressed
relative to cortisol and used to estimate effective minimum doses.
Preferred corticosteroids for use according to the invention include, but are
not limited
to, dexamethasone, prednisone, prednisolone, hoc-methylprednisolone,
fludrocortisone,
triamcinolone, paramethasone, betamethasone, aldosterone, and pharmaceutically-
acceptable
salts thereof. The corticosteroids may be administered according to the
invention as solvates
thereof or in the form of a pharmaceutically acceptable salt or ester.
Suitable salts or esters
include the acetate, isonicotinoate; phenylpropionate, pivalate, t-butyl
acetate,
to trioxaundecanoate, disodium metasulphobenzoate and disodiom phosphate.
For the present invention, dexamethasone is demonstrated as one example of a
synthetic corticosteroid with the ability to alleviate stress in an animal,
specifically transport
stress. Since corticosteroids essentially share the same functions but to
varying degrees, it
will be appreciated by those skilled in the art that alternative natural or
synthetic
corticosteroids similar in structure and function to dexamethasone may be
appropriate for the
present invention. Further, use of more than one corticosteroid in combination
or as a
"cocktail" might have appreciable effects by "balancing" the different actions
of each
corticosteroid.
While the invention is demonstrated as particularly useful for transport
stress, it will
be appreciated by those skilled in the art that the invention is equally
suitable for other
stressors involving high-energy demands over prolonged periods and preceded by
stressors
that initiate HPA axis response. Such types of stress include, but are not
limited to,
transportation stress, antemortem stress, regrouping stress, handling and
management stress,
such as moving, herding, walking, branding, neutering, dehorning or antler
removal; mixing
of animals, post-parturient stress, particularly as applicable to the
transition dairy cow;
weaning stress; acute weather stressors; noise stressors, particularly acute
noise stressors; and
on occasions when a non-steady physiological state is being experienced by an
animal.
As endogenous corticosteroid response to high-energy demand stressors is
similar
among all vertebrates, the invention can be applied to a variety of animal
species, particularly
8
CA 02470417 2004-06-09
non-human animals including, but not limited to, domestic ruminant and
monogastric
animals, including swine (Sus domesticus), horses, cattle (Bo~s taurus and Bos
indicus);
domestic ungulates, including bison, sheep, lamb, deer, moose, elk, caribou
and goats;
domesticated fowl, including chickens, turkeys, geese, ducks, game birds, and
other birds
raised in domestication to produce eggs or meat; dogs, cats and other
companion animals;
wild animals; captured animals; and zoo animals.
I. FORMULATIONS, DOSAGES AND ADMINISTRATION
io a) Corticosteroid Component
Various formulations of the corticosteroid are ideal for administration to
animals to
mitigate the effects of stress. The corticosteroid can be formulated as a
solid, liquid,
suspension, aerosol, liquid injectable, topical preparation, feed additive,
admixture, and feed
composition as follows.
i) Solids
The corticosteroid can be formulated as a solid, capsule, crumble, granule,
pellet, pill,
powder, tablet and similar dosage form. Corticosteroids may be applied
directly into feed
bunks or mixed with a ration.
ii) Liquids> Suspensions
The corticosteroid can be incorporated into liquids, formulated as solutions
or
suspensions by adding powdered corticosteroid to a suitable liquid. A
corticosteroid can be
mixed with the animal's drinking water or provided in other liquid forms for
consumption.
iii) Aerosols
Solutions of the corticosteroid can be converted into aerosols or sprays by
standard
techniques for making aerosol pharmaceuticals. In general, such techniques
comprise
pressurizing or providing a means for pressurizing a container of the
solution, usually with an
inert earner gas, and passing the pressurized gas through a small orifice.
Aerosols can
additionally contain customary propellants, for example, inert gases such as
nitrogen, carbon
dioxide, argon or neon.
9
CA 02470417 2004-06-09
iv) Liquid Injectables
The corticosteroid can be incorporated into a sterile injectable solution by
adding the
corticosteroid in the required amount in an appropriate pharmaceutically- or
therapeutically-
acceptable carrier (for example, physiological saline or other suitable
carriers) followed by
filtration, sterilization, or other techniques which ensure that the liquid
injectable meets
sterility and pyrogenicity requirements for veterinary use. Liquid solutions
and formulations
of the corticosteroid may lose some activity with aging and are thus either
prepared in stable
forms, preferably prepared fresh for administration, or include stabilizers,
solubilizers and
preservatives as are well known, see for example, Goodman and Gilman (1990).
v) Topical Preparations
Topical pharmaceutical compositions, for example creams, lotions, gels, pastes
and
ointments, and other topical forms such as liposomes, skin patches, etc. can
be used for
transcutaneous delivery of the corticosteroid to the body.
vi) Feed Additive
The corticosteroid can be administered in the form of a feed additive. The
feed
additive may be included with the animals' regular feed. Suitable feeds
include, but are not
limited to, grasses, legumes, grains, oil seeds, forbes, and sedges, for
example oats, barley,
wheat, canola, rye, sorghum, millet, corn, molasses, alfalfa, clover, brome,
timothy or fescue,
bermuda grass, orchard grass, rice straw, and other suitable feedstuffs. A
feed additive may
comprise the corticosteroid in combination with one or more inert or active
ingredients.
vii) Admixture
Incorporation of active ingredients into feed material is commonly achieved by
preparing a premix of the active ingredient, mixing the premix: with vitamins
and minerals,
and then adding the premix or feed additive to the feed. The corticosteroid
can be admixed
with other active ingredients known to those in the art. The active
ingredients, including the
corticosteroid alone or in combination with other active ingredients, can be
combined with
nutrients to provide a premixed supplement. Nutrients includes both
micronutrients, such as
vitamins, minerals, and macronutrients. The premix may then be added to feed
materials.
CA 02470417 2004-06-09
viii) Feed Composition
The corticosteroid can be provided in the form of a feed composition
comprising a
feed treated with the corticosteroid. The corticosteroid may lbe mixed with a
feed in dry form;
e.g. as a powder, or as a liquid to be used as a drench or spray for example.
Suitable feeds
include, but are not limited to, grasses, legumes, grains, oil seeds, forbes,
and sedges, for
example oats, barley, wheat, canola, rye, sorghum, millet, corn, molasses,
alfalfa, clover,
brome, timothy or fescue, Bermuda grass, orchard grass, rice straw, and other
suitable
feedstuffs.
l0 Typically, the corticosteroid will be formulated in one or more of the
forms set out
above. These formulations may be stabilized through the addition of proteins
or chemical
agents. The corticosteroid can be prepared alone or as an active ingredient in
pharmaceutical
compositions including non-toxic, pharmaceutically or therapeutically
acceptable carriers,
diluents, excipients, antibiotics, prebiotics, probiotics, micronutrients,
vitamins, minerals, and
macronutrients, as are well known, see for example, Merck (1973) and Haynes
(1990). For
standard dosages of conventional pharmacological agents, see for example,
Merck (1973).
To ensure that the animal consumes a sufficient quantity, flavorings may be
added to provide
the corticosteroid in a form which appears palatable to the animal. All agents
must be non-
toxic and pharmaceutically-acceptable for the intended purpose, and must not
substantially
2o interfere with the effect of the corticosteroid.
The dosage of the corticosteroid depends upon many factors that are generally
known
to those skilled in the art, for example, the species, age, and weight of the
animal; the choice
of corticosteroid and its potency; the type and severity of stress; the time
and route of
administration; and the type and concentration of the formulation being
applied. Appropriate
amounts in any given instance will be readily apparent to those skilled in the
art or capable of
determination by routine experimentation. A therapeutically effective amount
of the
corticosteroid is desired, namely any amount of a formulation of the
corticosteroid which will
alleviate the effects of stress when administered to the animal in need
thereof. In general, the
3o minimum dose required to mediate a stress modulating effect is the critical
factor, and can be
estimated from the relative potencies according to Haynes (1990). The maximum
dose can be
taken as approximately ten times the minimum dose.
11
CA 02470417 2004-06-09
The corticosteroid can be administered prior to exposure to the stress, during
the
stress, andlor following exposure to the stress. Moreover, the corticosteroid
can be
administered in several ways, including, for example, orally, intramuscularly,
intravenously,
infra-nasally, via suppository, or transcutaneously, with orally being the
most convenient
method.
When administered orally (e.g., as a food or water additive), the
corticosteroid may be
provided, for a 500 kg animal, at a dosage range of 1 - 20 mg, more preferably
2 - 5 mg , and
most preferably 1 - 5 mg. For transport stress, oral forms can be administered
within 0 - 48
to hours prior to transport, more preferably within 6 - 12 hours prior to
transport, and most
preferably within 0 - 6 hours prior to transport. A longer acting
corticosteroid may be
administered within 12 - 24 hours prior to transport.
When administered intramuscularly, the corticosteroid may be provided, for a
500 kg
animal, at a dosage range of l - 20 mg, more preferably 2 - 5 ~mg, and most
preferably 1 - 5
mg. For transport stress, intramuscular injection of the corticosteroid is
conducted within 0 to
48 hours prior to transport, more preferably within 6-12 hours prior to
transport, and most
preferably 0 - 6 hours prior to transport. Table 1 provides an example of a
dosage schedule
for intramuscular administration of common corticosteroids prior to transport.
The doses
2o apply to cattle of approximately 1000 lb bodyweight. The indicated
"transport time" refers
to the transport period for which the particular corticosteroid is beneficial.
12
CA 02470417 2004-06-09
Table 1. Dosage Schedule for Intramuscular Administration of Corticosteroids
Prior to
Transport or Specific Transport Times
Corticosteroid Minimum Transport Time (hr)
intramuscular Dose
(mg)
Dexamethasone 10 0 - 48
Prednisone 50 0 - 24
Prednisolone 50 0 - 24
Got-methylprednisolone50 0 - 24
Fludrocortisone 20 0 - 12
Triamcinolone 40 0 - 24
Paramethasone 20 0 - 48
Betamethasone 10 0 - 48
Aldosterone 10 0 - 24
When administered intravenously, the corticosteroid may be provided, for a 500
kg
animal, at a dosage range of 1 - 20 mg, more preferably of 2 - S mg, and most
preferably of 1
- 5 mg. For transport stress, intravenous administration of the corticosteroid
is conducted
within 0 to 48 hours prior to transport, more preferably within 6-12 hours
prior to transport,
and most preferably 0 - 6 hours prior to transport. Further, the
corticosteroid may be
administered intravenously to an animal exhibiting severe signs of distress
immediately
following transport.
Further, the corticosteroid can be administered infra-nasally using, for
example, an
aerosol or spray, to deliver the corticosteroid to the animal via the mucosal
tissues. However,
effective infra-nasal delivery of the corticosteroid is dependent upon several
factors, for
example, the receptiveness of the animal to aerosol administration with
respect to the
anatomy and physiology of its nasal cavity, and accessibility of the olfactory
region. If intra-
nasal administration is unsuitable for small nasal cavities of particular
animals, a suppository
containing the corticosteroid can be used for placement into an appropriate
orifice of the
13
CA 02470417 2004-06-09
animal where it melts at body temperature, releasing the corticosteroid. A far
less invasive
technique involves transcutaneous administration. The corticosteroid, in
dosage forms
suitable for epidermal application (for example, topical preparations such as
creams, lotions,
gels, pastes, and ointments; liposomes; skin patches; etc.), is thereby
applied directly to the
skin of the animal for a systemic effect. The dosage ranges a.nd schedules as
previously
discussed for oral, intramuscular and intravenous administrations are likewise
appropriate for
intra-nasal, suppository, and transcutaneous administrations.
Approaches for sustaining or controlling release of the corticosteroid
include, for
example, microspheres, microcapsules, projectile biodegradable missiles and
the like. The
corticosteroid can be incorporated into a biodegradable polymer which can be
injected as a
microsphere, for example, reservoir devices (encapsulation of the
corticosteroid within a
polymer shell) or matrix devices (the corticosteroid is physically entrapped
within a polymer
network). In addition, implantable controlled-release drug delivery systems,
for example,
I5 pellets (composed of various polymeric forms), compressed tablets, silastic
rubber implants,
and silicone capsules, are suitable. The corticosteroid can be incorporated
into a
biodegradable polymer and the mixture shaped into a disc, fiber or other form
for
implantation. The implant can then be inserted into the animal through an
incision.
Examples of implantable devices for sustained or controlled delivery of the
corticosteroid
within the animal include osmotically or propellant-driven pumps, infusion
pumps, mini-
pumps, and the like.
b) Corticosteroid in Conjunction with a Nutritional Supplement
Surprisingly, the inventors have found that optimal synergistic effects are
obtained
when the corticosteroid is used in conjunction with a nutritional supplement.
The nutritional
supplement for use in the present invention is described in United States
Patent Nos.
5,505,968 and 5,728,675 to Schaefer et al. issued April 9, 1996 and March 17
respectively, of
which the teachings are applicable to the present invention and are
incorporated by reference
herein. The nutritional supplement provides the following effects or benefits
from the
3o individual ingredients:
i) The electrolyte imbalance from stress is corrected and/or normalized by the
inclusion
of Na, K and Mg, and preferably bicarbonate.
14
CA 02470417 2004-06-09
ii) The hypoglycaemic condition that arises from stress is corrected and/or
normalized by
the inclusion of a source of energy, preferably glucose, together with the
gluconeogenic precursor alanine.
iii) The dehydration associated with stress is corrected andlor normalized
either by the
inclusion of water in the supplement itself (in the liquid forms of the
supplement) or
by the provision of water in conjunction with the supplement (in the solid
forms of the
supplement).
iv) Net protein degradation and carcass loss arising from stress is attenuated
by the
provision of specific amino acids including leucine, is~oleucine and valine,
which
to stimulate protein synthesis and reduce protein degradation, together with
the essential
amino acids including phenylalanine, lysine, threonine and methionine needed
in
protein synthesis. For ruminant animals, the amino acids are provided in a
bypass
form to ensure that they can be utilized by specifically ruminant animals and
to ensure
that there is a prolonged effect from the supplement after administration to
the animal.
IS v) Hypertension and anxiety experienced from stress is lessened by
including the amino
acid tryptophan, which is the neurotransmitter precursor to se:rotonin,
together with
the blood pressure lowering agent magnesium sulphate. This amino acid is
provided
in a bypass form for ruminants and/or food grade form.
vi) The effects of meat quality degradation from stress are also lessened by
the combined
20 action of electrolytes, which promote acid/base stability and buffering,
with NH3
recipients (glutamate). Protein degradation in animals results in the release
of NH3
groups, which can lead to high pH conditions known to contribute to dark
cutting
problems in meat quality. The provision of glutarnic acid (glutamate) in the
supplement alleviates the NH3 buildup problem. The glutamate, or glutamic
acid, is
25 preferably provided in relatively large concentrations, compared to that of
other amino
acids, to provide an ammonia buffering effect.
Briefly, the nutritional supplement contains an energy ;>ource, preferably
glucose, up
to 1000 g per 500 kg animal. The source of energy is met witri the inclusion
of simple or
30 complex carbohydrates or fats. Preferable energy sources include one or
more of glucose,
sucrose, fructose, galactose, dextrose, propylene glycol, lactose, complex
carbohydrates such
as starch, and fat. Several of these ingredients are beneficially included to
delay the effect of
CA 02470417 2004-06-09
the energy source. For instance lactose, starch, propylene glycol, sucrose and
fat provide
prolonged energy sources. The energy source is preferably provided in a form
which is
palatable and familiar to the animal. Such sources as kraft whey powder,
molasses, and skim
milk powder are economic forms of energy sources which are particularly
preferred alone, or
in admixture with purer energy sources such as glucose, sucrose and dextrose.
Other useful
energy sources will be evident to persons skilled in the art.
The nutritional supplement also provides Na, K and Mg, preferably at a
hypotonic
concentration. Ion complexes such as NaCI, KHC03 and MgSO~ are preferred and
provide
10 to 20 g of actual ingredient per 500 kg animal.
A source of specific amino acids is provided, including alanine, lysine,
phenylalanine,
methionine, threonine, leucine, isoleucine, valine and tryptophan at a minimum
of 0.5 g per
500 kg animal. Leucine additions at 15 g and glutamate at 40~ g per 500 kg
animal are
preferred. For ruminants, amino acids are provided in a bypass form. The amino
acid
complex was designed to stimulate protein synthesis, counter protein
catabolism and
contained the neurotransmitter precursors tryptophan and tyrosine, designed to
attenuate the
HPA axis response.
The respective amounts of the individual ingredients vary from animal to
animal.
Generally, as a guide, based on a body weight/three quarters power scale, for
a 500 kg .
ruminant animal consuming about 20 L of water in a day, the ;preferred ranges
of ingredients
are as follows (g ingredient/500 kg animal.):
Energy source (based on glucose) 20-2000 g (pref. 50-1000)
Electrolytes (NaCI, KHC03, each) 2-40 g (pref. 10-20)
(MgS04) 1-20 g (pref. 10)
Amino Acids (all) 0.5-100 g (pref. 2-10)
(leucine) 15-25 g
(glutamic acid) 40-66 g
16
CA 02470417 2004-06-09
Exemplary ingredients (if present) are most preferably included in the
following
percent by weight amounts:
Feed Grade ingredients
(preferred range is 0.1-4 times the amounts set out below:
Flavour 1 %
Methionine 0.5% (as pure source)
Lysine 0.3% (as pure source)
Tryptophan 0.4-l % (as pure source)
Threonine 0.15% (as pure source)
Magnesium sulphate 1-2% (as epsom salts)
' Potassium chloride l .5%
Sodium chloride 2-4%
Potassium bicarbonate 4%
Sodium bicarbonate 4%
Dextrose 20%
Animal or Vegetable 2%
fat
Molasses 4%
Sources of Lactose:
Skim milk powder 2.5-15%
Whey powder 10-20%
17
CA 02470417 2004-06-09
Bypass Proteins as Sources of Amino Acids:
Cotton seed meal 15-50%
Corn gluten meal 15-40%
Distillers grain 30-60°Io
Hydrolysed feather meal 10-20%
Fishmeal 20-30%
Meat and bone meal 20-40%
Blood meal 10-20%
Various formulations of the nutritional supplement are fully described in
United
States Patent Nos. 5,505,968 and 5,728,675 to Schaefer et al., for example,
solid feed
supplements (preferably as a pelletized solid for admixture with the normal
food for the
animal), powder premixes for dilution into liquid for either drench or liquid
consumable
products, or as concentrated liquids for drenches or liquid consumables (with
or without
dilution). The supplement is most preferably administered as a preventative
nutrient
supplement before the animal is exposed to the stress. For transport stress,
the supplement is
preferably administered 6-24 hours prior to transport and most preferably 6-12
hours prior to
transport.
Examples 1 and 2 illustrate the effects of corticosteroid alone, and
corticosteroid in
conjunction with a nutritional supplement upon weight toss normally incurred
during
transport and handling stress. In Example 1, two treatment groups designated
as control, and
dexamethasone-treated were studied. In Example 2, three groups designated as
control,
dexamethasone-treated and a nutritional supplement plus dexa.methasone were
studied. The
results indicate that a corticosteroid, such as dexamethasone, alone and in
conjunction with a
nutritional supplement is effective in reducing weight loss due to transport
and handling
stress. Example 3 demonstrates that corticosteroid treatment in conjunction
with a nutritional
18
CA 02470417 2004-06-09
supplement appears to enhance recovery from transport stress, in that animals
displayed
behavioral changes, namely, an improvement in appetite, and a decline in need
to repose.
Overall, Examples l, 2 and 3 suggest that a synthetic corticosteroid, having a
potency and
biological half-life greater than that of endogenous corticosteroid, can
augment the HPA axis
response and assist animals in maintaining homeostasis and coping physically
with stress.
The invention is illustrated in the following non-limiting examples.
Example 1- Effects of Corticosteroid Treatment
Forty head of crossbred calves, both steers and heifers, and weighing
approximately
600 lb (272 kg) on average were used. The calves were raised at the
Agriculture and Agri-
Food Canada Lacombe Research Centre (Lacombe, Alberta, Canada) in accordance
with
standard operating procedures representative of the cattle industry. The
calves were randomly
divided into two treatment groups designated as control (n=20) and
dexamethasone-treated
(n=20) and kept on standard feed (cereal grain silage) and water rations prior
to transport.
Weights of all animals were recorded pre- and post-transport, metabolic
activity was
assessed by infrared thermographic analyses (1RT) of the eye region, and a
salivary swab
collection made for cortisol measurement. The dexamethasone-treated calves
were then
given an intramuscular dose of dexamethasone at 2 mg per 50 lb (23 kg) body
weight.
The animals were subsequently loaded onto a commercial transport and
transported
for l Oh prior to being off loaded at the Agriculture and Agri-Food Canada
Kamloops
Research Centre (Kamloops, British Columbia, Canada). On arrival, the calves
were again
weighed, infrared images captured and a salivary swab collected. The animals
were held
overnight in pens with ad libitum access to water and cereal grain silage, and
were again
monitored for weight, thermal changes and salivary cortisol the following
morning.
The IRT temperature was significantly lower for the dexamethasone-treated
group
than the control group following transport (P<0.04). The use of dexamethasone
thus results
in a lowered thermal response in animals.
19
CA 02470417 2004-06-09
For the period of transport between Lacombe and Kamloops (10 h), the control
calves
lost on average 30.5 lb (13.8 kg or 5% body weight), while the dexamethasone-
treated calves
lost 19.2 lb (8.7 kg or 3.2% body weight) by comparison. This difference in
weight loss was
statistically significant (P<0.05). The results suggest that dexamethasone
treatment
demonstrated the potential to significantly reduce weight loss in transported
cattle.
Example 2 - Effects of Corticosteroid Treatment in Conjunction with a
Nutritional
Supplement
The study in Example 1 was repeated with the addition of a nutritional
supplement
to treatment group. The calves were raised at the Agriculture and Agri-Food
Canada Lacombe
Research Centre (Lacombe, Alberta, Canada) in accordance with standard
operating
procedures representative of the cattle industry. The breed, sex and weight of
the calves were
similar to those used in Example 1. Crossbred calves weighing approximately
800 lb (363
kg) on average were used.
Three treatment groups were designated as control (n==15), dexamethasone-
treated or
DEX (n=15) and dexamethasone plus nutritional supplement or DEX + NT (n=15).
Twenty-
four hours prior to transport, the calves were assigned to the nutritional
supplement groups
and were offered 1 kg/animal of nutritional supplement or a "receiver calf'
preparation
(Supplement 1 (d) as described in United States Patent Nos. 5,505,968 and
5,728,675 to
Schaefer et al. with differences being the selection of flavoring, form of an
extruded pellet,
and carrier being alfalfa) along with their regular feed ration (cereal grain
silage). All calves
in all treatments were kept on normal feed and water rations prior to
transport.
Weights of all animals were recorded pre- and post-transport and metabolic
activity
was assessed by infrared thermographic analyses of the eye region. The morning
of transport
all animals were weighed, scanned with infrared cameras and had a salivary
swab collected.
The dexamethasone treatment given just prior to transport consisted of an
intramuscular
injection of 0.61 mg per 50 lb body weight. The animals were then loaded onto
a commercial
transport earner and transported for 8h prior to being offloaded, re-weighed,
re-scanned with
infrared and re-sampled for cortisol. For the DEX -~ NT animals, the calves
were provided on
CA 02470417 2004-06-09
arrival, a further lkg/head of a liquid tank mixable and 1 kg/head "receiver
calf' pellets in
addition to the cereal silage.
The control animals demonstrated a significant increase in temperature over
the
transport period (P<0.01). Treated animals also exhibited a rise in IIZT
temperature but to a
significantly smaller degree compared to the controls (P<0.05). The use of
dexamethasone,
or dexamethasone plus nutritional supplement thus results in a lowered thermal
response in
treated animals.
to The control animals lost on average 47.36 lb (21.5 kg or 5.6% body weight)
during
transport compared to a loss of 38.87 lb (17.6 kg) for the dexamethasone
treated calves and a
loss of 30.2 lb (13.7 kg or 3.4°lo body weight) for the combined
treatment group of
dexamethasone and nutritional supplement. These differences were significantly
different at
P<0.05. The results indicate that treatment with corticosteroids and with
corticosteroids in
conjunction with a nutritional supplement resulted in significantly less
weight loss in
transported cattle. Overall, in both Examples 1 and 2, DEX or DEX + NT groups
exhibited
significantly less weight losses than Controls (P<0.01 ). The effects of the
nutritional
supplement and corticosteroids were synergistic in alleviating transport and
handling stress in
cattle.
Example 3 - Effects of Corticosteroid Treatment in Conjunction wvith a
Nutritional
Supplement on Recovery
Corticosteroid treatment in conjunction with a nutritional supplement appears
to
enhance recovery from transport stress. The behavioral responses of animals
were monitored
following transport. Calves were allocated among five treatment groups as
follows:
21
CA 02470417 2004-06-09
Table 2. Treatment Groups to Examine Effects of Corticosteroid Treatment in
r'nninnctinn with a Nutritional Sunnlement on Recoyery
Group # of Description
Animals
Control home (CH) I6 animals stay~:d in a pen at
the Lacombe
Research Centre and were not
exposed to
transport
Control away (CA) 15 control animals exposed to the
same
transport conditions as the
treatment calves
Nutritional supplement15 animals treated with a nutritional
(NT)
supplement prior to transport
Nutritional supplementI S animals treated with a nutritional
plus
dexamethasone (NT supplement plus dexamethasone
+ DEX)
Dexamethasone alone 15 animals treated with dexamethasone
(DEX) alone
For all of the above groups; ten behavioural scans lasting one minute each
were
completed each day (pm) for three days following transport, thereby
accumulating a total of
2,280 animal observations. Frequency measures of animals lying and eating were
collected.
It is well known to those skilled in the art that when animals ace fatigued or
morbid, they tend
to lie down (Kilbour et al., 1984; Agri-Food Research Council Canadian Food
Inspection
Agency, 2001). Further, animals that are morbid display a reduced feeding
frequency,
to whereas healthy, non-fatigued and hungry animals exhibit an increased
feeding frequency.
The CA animals displayed a three day average of 97% lying compared to only 23%
in
the CH calves. In contrast, the NT calves displayed a 45% lying frequency; the
NT + DEX a
39% frequency and the DEX a 0% lying frequency. Such behavioural data suggest
that the
NT, NT + DEX, and DEX animals ali displayed an improved ,recovery rate based
on their
reduced need to lie down.
The eating frequency data also supported this conclusion with the three day
frequency
average for the CA calves at 20% compared to 43%, 35% and 24% in the NT, NT +
DEX and
2o DEX animals respectively. In summary, the frequency behaviour results
suggest that the
DEX, and DEX + NT treated animals recovered faster than controls.
22
CA 02470417 2004-06-09
Table 3. Results of Corticosteroid Treatment in Conjunction with a Nutritional
Supplement on Recovery
Group Lying Frequency Eating Frequency
( % ) ( % )
Control home (CH) 23 78
Control away (CA) 97 20
Nutritional supplement45 43
(NT)
Nutritional supplement39 35
plus
dexamethasone (NT
+ DEX)
Dexamethasone alone 0 - ~ 24
(DEX)
23
CA 02470417 2004-06-09
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All publications mentioned in this specification are indicative of the level
of skill of
those skilled in the art to which this invention pertains. All publications
are herein
incorporated by reference to the same extent as if each individual publication
was specifically
and individually indicated to be incorporated by reference.
The terms and expressions in this specification are, unl!,ess otherwise
specifically
3o defined herein, used as terms of description and not of limitation. There
is no intention, in
using such terms and expressions, of excluding equivalents of the features
illustrated and
26
CA 02470417 2004-06-09
described, it being recognized that the scope of the invention is defined and
limited only by
the claims which follow.
27
