U.S. patent application number 14/465634 was filed with the patent office on 2014-12-25 for methods of making and using a ruminant gas reduction composition.
The applicant listed for this patent is Georgia-Pacific LLC. Invention is credited to Anne Chace Hopkins, Thomas A. Lehtinen, Matthew W. Lowe.
Application Number | 20140378412 14/465634 |
Document ID | / |
Family ID | 43733048 |
Filed Date | 2014-12-25 |
United States Patent
Application |
20140378412 |
Kind Code |
A1 |
Lowe; Matthew W. ; et
al. |
December 25, 2014 |
METHODS OF MAKING AND USING A RUMINANT GAS REDUCTION
COMPOSITION
Abstract
A method comprising administering an oligosaccharide composition
to an organism having a gastrointestinal system to affect the
production of GHG produced by the organism allowing for a reduction
of the GHG produced by the organism while optimizing the health,
feed intake, and protein synthesis of the organism so that
management of the organism may realize the synergistic effects of
maximizing both typical organism commodity-type concerns (e.g.,
size and production metrics) and atypical organism commodity-type
concerns (e.g., carbon credit trading/monetization). A gas-reducing
composition comprising soluble extractable material from a
lignocellulosic source. A method of producing a composition,
comprising providing a lignocellulosic source; extracting soluble
materials from the lignocellulosic source to produce soluble
extractable material; and processing the soluble extractable
material to yield a gas-reducing composition, wherein the
gas-reducing composition comprises hemicellulose and exhibits
gas-reducing activity.
Inventors: |
Lowe; Matthew W.; (Lufkin,
TX) ; Hopkins; Anne Chace; (Diboll, TX) ;
Lehtinen; Thomas A.; (Diboll, TX) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Georgia-Pacific LLC |
Atlanta |
GA |
US |
|
|
Family ID: |
43733048 |
Appl. No.: |
14/465634 |
Filed: |
August 21, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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13392288 |
Feb 24, 2012 |
8828970 |
|
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PCT/US2010/046867 |
Aug 26, 2010 |
|
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14465634 |
|
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|
61237396 |
Aug 27, 2009 |
|
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Current U.S.
Class: |
514/54 ;
536/123.1 |
Current CPC
Class: |
A61P 1/00 20180101; A61K
31/715 20130101; A23K 20/163 20160501; A23K 50/10 20160501 |
Class at
Publication: |
514/54 ;
536/123.1 |
International
Class: |
A61K 31/715 20060101
A61K031/715 |
Claims
1. A ruminal gas reducing composition comprising soluble
extractable material from a lignocellulosic source.
2. The composition of claim 1, wherein the soluble extractable
material comprises galactoglucomannans, xylans, fructans,
arabinoxylans, glucomannans, derivatives thereof, or combinations
thereof.
3. The composition of claim 1, wherein the soluble extractable
material comprises galactoglucomannans.
4. The composition of claim 3, wherein the galactoglucomannans
comprise glucose units, galactose units, and mannose units in a
ratio of about 3 to about 1 to about 6.
5. The composition of claim 1, wherein the lignocellulosic source
comprises a member of the order Pinales, the family Pinaceae, a
member of the family Fagaceae, a member of the order Saxifragales,
or combinations thereof.
6. The composition of claim 5, wherein the lignocellulosic source
comprises a member of the genus Pinus.
7. A method of producing a composition, comprising: providing a
lignocellulosic source; extracting soluble materials from the
lignocellulosic source to produce soluble extractable material; and
processing the soluble extractable material to yield a gas-reducing
composition, wherein the gas-reducing composition comprises
hemicellulose and exhibits gas-reducing activity.
8. The method of claim 7, wherein extracting soluble materials
comprises softening the lignocellulosic source by autohydrolysis,
pulping, steam explosion, steam extrusion, or combinations
thereof.
9. The method of claim 7, wherein the hemicellulose comprises
galactoglucomannans, xylans, fructans, arabinoxylans, glucomannans,
derivatives thereof, or combinations thereof.
10. The method of claim 7, wherein the soluble extractable
materials comprise monosaccharides, oligosaccharides, and
polysaccharides composed of glucose, galactose, and mannose units
in a ratio of about 3 to about 1 to about 6.
11. The method of claim 7 further comprising hydrolyzing the
soluble extractable materials to produce a hydrolyzed
composition.
12. The method of claim 11, wherein the hydrolyzed composition
comprises polysaccharides having a degree of polymerization of from
about 2 to about 20.
13. The method of claim 7 further comprising dehydrating the
soluble extractable materials.
14. A method comprising administering the gas-reducing composition
of claim 1 to an organism having a gastrointestinal system
including an organism that has a rumen in its gastrointestinal
tract.
15. The method of claim 14, wherein administration of the
gas-reducing composition results in a decrease in the methane
production of the organism when compared to an otherwise similar
organism not administered the gas-reducing composition.
16. The method of claim 14, wherein administration of the
gas-reducing composition results in an increase in the dry matter
digestibility when compared to an otherwise similar organism not
administered the gas-reducing composition.
17. The method of claim 14, wherein administration of the
gas-reducing composition results in an change in volatile fatty
acid production, a change in pH, and an decreased
acetate:propionate ratio as compared to an otherwise similar
organism not administered the gas-reducing composition.
18. The method of claim 14, wherein administration of the
gas-reducing composition results in an increase in hexose and/or
pentose fermentation within the organism when compared to an
otherwise similar organism not administered the gas-reducing
composition.
19. The method of claim 14, wherein administration of the
composition improves production of an organism-derived commodity, a
biological function, or combinations thereof.
20. The method of claim 19, wherein the biological function
comprises nutrient uptake, muscle growth, muscle development,
weight gain, coat growth, survival, or combinations thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of and claims priority to
U.S. patent application Ser. No. 13/392,288 filed on Feb. 24, 2012,
published as US 2012/0225841 A1, which is a filing under 35 U.S.C.
371 of International Application No. PCT/US2010/046867 filed Aug.
26, 2010, entitled "Methods of Making and Using a Ruminant Gas
Reduction Composition," claiming priority of U.S. Provisional
Patent Application No. 61/237,396 filed on Aug. 27, 2009, which
applications are incorporated by reference herein in their
entirety.
BACKGROUND
[0002] Greenhouse gases ("GHG") are gases in an atmosphere that
absorb and emit radiation within the thermal infrared range. The
primary greenhouse gases in the Earth's atmosphere are water vapor,
carbon dioxide, methane, nitrous oxide, and ozone.
[0003] The 2010 United States federal budget proposes to support
clean energy development with a 10-year investment of $15 billion
per year, generated from the sale of GHG emissions credits. Under
the proposed cap-and-trade program, all GHG emissions credits would
be auctioned off, generating an estimated $78.7 billion in
additional revenue in FY 2012, steadily increasing to $83 billion
by FY 2019.
[0004] Emissions trading is a market-based approach used to control
pollution by providing economic incentives for achieving reductions
in the emissions of pollutants. Governing entities may establish a
limit or cap on the amount of a pollutant that can be emitted. Such
limit or cap may be applied, allocated, or sold to entities which
have been identified as capable of producing emissions at a level
which could be subject to the established limit or cap for said
designated pollutants. These limits or caps may be applied,
allocated, or sold to such emissions entities in the form of
emissions permits which represent the right to emit or discharge a
specific volume of a specified pollutant. Such emission producing
entities are required to hold a number of permits (or credits)
equivalent to their emissions. The total amount of permits (or
credits) issued by the governing entity cannot exceed the cap;
thus, limiting total emissions to that level. Emissions entities
that need to increase their level of emissions must buy permits
from those who require fewer permits. The transfer of permits is
referred to as a trade. In effect, the buyer is paying a charge for
polluting, while the seller is being rewarded for having reduced
emissions.
[0005] The overall goal of an emissions trading plan is to minimize
the cost of meeting a set emissions target. The cap is an
enforceable limit on emissions that is usually lowered over
time--aiming towards a national emissions reduction target. In
other systems a portion of all traded credits must be retired,
causing a net reduction in emissions each time a trade occurs.
Thus, in theory, by limiting or capping polluting emissions the
totality of pollution may be decreased. Moreover, those who can
reduce emissions most cheaply will do so, achieving pollution
reduction at the lowest cost to society.
[0006] There are active trading programs in several air pollutants.
For GHG the largest is the European Union Emission Trading Scheme.
In the United States there is a national market to reduce acid rain
and several regional markets in nitrogen oxides. In 2003, New York
State proposed and attained commitments from nine Northeast states
to form a cap-and-trade carbon dioxide emissions program for power
generators, called the Regional Greenhouse Gas Initiative. This
program launched on Jan. 1, 2009 with the aim to reduce the carbon
"budget" of each state's electricity generation sector to 10% below
their 2009 allowances by 2018. Also in 2003, U.S. corporations were
able to trade CO.sub.2 emission allowances on the Chicago Climate
Exchange under a voluntary scheme. In August 2007, the Exchange
announced a mechanism to create emission offsets for projects
within the United States that cleanly destroy ozone-depleting
substances.
[0007] Since February 2007, seven U.S. states and four Canadian
provinces have joined together to create the Western Climate
Initiative, a regional GHG emissions trading system. July 2010, a
meeting took place to further outlined the cap-and-trade system
which if accepted would curb GHG emissions by January 2012.
[0008] In 2006, the California Legislature passed the California
Global Warming Solutions Act, AB-32. Project based offsets have
been suggested for five main project types. A carbon project would
create offsets by showing that it has reduced carbon dioxide and
equivalent gases. The project types include: building energy,
landfill gas capture, forestry, and manure management.
[0009] According to Food and Agriculture Organization statistics,
ruminant livestock-derived methane has been estimated at 18% of the
total global GHG emissions on a carbon dioxide equivalency basis.
In addition, global protein consumption more than doubled since
1970 and is projected to double again by 2050. Ruminant-derived
methane is produced during digestion (fermentation) of feed and
fodder through microbial fermentation within the rumen. Ruminant
methane levels are attributable to the rate, efficiency, and
completeness of carbohydrate and protein conversion from feedstuffs
into volatile fatty acids ("VFAs"). The molar percentage and
composition of ruminal VFAs produced during fermentation influence
the production of methane. Acetate and butyrate promote methane
production while propionate formation is considered a competitive
pathway for hydrogen use in the rumen.
[0010] There is an inverse relationship between fermentation
efficiency and methane production within the rumen. Metabolic
energy loss during rumen digestion can be attributed to heat loss
during fermentation, as well as the production of ammonia and
methane gas. Methane reduction within the rumen not only improves
GHG emissions but is attributable to increased energy conversion
and subsequent enhanced animal productivity. These benefits are of
key interest to farmers and producers.
[0011] Feed (diet) and feeding strategies have demonstrated
significant influence on fermentation products and energy
production within ruminant animals. Rumen bypass protein products
are an exceptional example of how nutritional manipulation can
benefit animal productivity. Methane reducing feed additives, many
of which are plant-based, however, have shown limited holistic
success, or have demonstrated adverse trade-offs that have
precluded their widespread practical application. These negative
effects include reduced feed intake and protein synthesis, both of
which can limit optimal growth and development. Natural feed
additives which could improve dry matter digestion and reduce
methane production would represent an appealing solution for
reducing livestock-derived GHG while contributing to optimal animal
nutrition. Thus, the natural feed additives described herein may
allow farmers and producers to maximize their food commodity
production, benefit from emissions trading programs, comply with
greenhouse gas emission mandates, regulations, and contribute to a
better global environment.
SUMMARY
[0012] Disclosed herein is an oligosaccharide composition
comprising soluble extractable material from a lignocellulosic
source wherein the soluble extractable material comprises a
hemicellulose. In an embodiment, the soluble extractable material
comprises galactoglucomannans, xylans, arabinoxylans, or
combinations thereof. In another embodiment the soluble extractable
material comprises galactoglucomannans and the galactoglucomannans
comprise glucose monosaccharide units, galactose monosaccharide
units, and mannose monosaccharide units in a ratio of about 3 to
about 1 to about 6. In an embodiment, the lignocellulosic source
comprises the above and below-ground portion of a plant wherein the
above-ground portion of a plant exhibits cambial growth. In another
embodiment, the lignocellulosic source comprises a member of the
family Pinaceae, a member of the family Fagaceae, a member of the
order Saxifragales, a member of the order Pinales, or combinations
thereof. In yet another embodiment, the lignocellulosic source
comprises a member of the genus Pinus. In another embodiment an
admixture comprises the oligosaccharide composition and one or more
pharmaceutical carriers.
[0013] Also disclosed herein is a method comprising administering
the oligosaccharide composition to an organism to reduce the
production of rumen-produced methane gas.
[0014] Also disclosed herein is a method comprising administering
the oligosaccharide composition to an organism to reduce the
production of ruminal ammonia.
[0015] Also disclosed herein is a feed product comprising the
oligosaccharide composition.
[0016] Also disclosed herein is an admixture of the oligosaccharide
composition with one or more feed products, feed liquids, feed
supplements, or combinations thereof.
[0017] Also disclosed herein is a method of producing a
composition, comprising a lignocellulosic source; extracting
soluble materials from the lignocellulosic source to produce
soluble extractable material; and processing the soluble
extractable material to yield a gas reducing composition, wherein
the composition comprises hemicellulose and exhibits the ability to
reduce methane and ammonia production in ruminants. In an
embodiment extracting soluble materials comprises softening the
lignocellulosic source. In an embodiment softening of the
lignocellulosic source comprises autohydrolysis, pulping, steam
explosion, steam extrusion, or combinations thereof. In an
embodiment the hemicellulose comprises monomers, oligosaccharides,
and polysaccharides having a degree of polymerization from 1 to
greater than about 500. In an embodiment the hemicellulose
comprises xylans, arabinoxylans, galactoglucomannans, manans,
derivatives thereof, or combinations thereof. In an embodiment the
soluble extractable materials comprise monosaccharides,
oligosaccharides, and polysaccharides composed of glucose,
galactose, and mannose units in a ratio of about 3 to about 1 to
about 6. In an embodiment, the method further comprises hydrolyzing
the soluble extractable materials to produce a hydrolyzed
composition. In an embodiment, the hydrolyzed composition comprises
polysaccharides having a degree of polymerization of from about 2
to about 20. In an embodiment, the method further comprises
dehydrating the soluble extractable materials.
[0018] Also disclosed herein is a method comprising administering
the oligosaccharide composition to an organism having a
gastrointestinal system. In an embodiment administration of the
oligosaccharide composition reduces the production of methane
and/or ammonia within the organism.
[0019] Also disclosed herein is a method comprising administering
the oligosaccharide composition to an organism having a
gastrointestinal system. In an embodiment administration of the
oligosaccharide composition reduces the production of ruminal
methane and/or ammonia within the organism.
[0020] Also disclosed herein is a method of managing livestock
comprising supplementing the livestock's diet with a gas reducing
composition comprising soluble extractable material from a
lignocellulosic source, quantifying, or having quantified, a
reduction in gas produced by the livestock subsequent to the
supplementation and realizing an economic or other benefit from the
reduction in gas produced.
[0021] Also disclosed herein is a method of managing livestock
comprising determining a baseline amount of greenhouse gases
produced by the livestock, administering to the livestock a gas
reducing composition comprising soluble extractable material from a
lignocellulosic source, determining an amount of greenhouse gases
produced by the livestock subsequent to administering the gas
reducing composition, calculating a reduction in greenhouse gases
by subtracting the amount of greenhouse gases produced by the
livestock subsequent to administering the gas reducing composition
to the baseline amount of greenhouse gases produced by the
livestock, and receiving an economic benefit from the reduction in
greenhouse gases produced by the livestock.
[0022] Also disclosed herein are a method and system comprising
administering the oligosaccharide composition to an organism having
a gastrointestinal system to affect the production of GHG produced
by the organism allowing for a reduction of the GHG produced by the
organism to be quantified and utilized in an emissions trading
program, for compliance with emission-related mandates, or to meet
the requirements of emission regulations.
[0023] Also disclosed herein are a method and system comprising
administering the oligosaccharide composition to an organism having
a gastrointestinal system to affect the production of GHG produced
by the organism allowing for a reduction of the GHG produced by the
organism while optimizing the health, feed intake, and protein
synthesis of the organism so that management of the organism may
realize the synergistic effects of maximizing both typical organism
commodity-type concerns (e.g., size and production metrics) and
atypical organism commodity-type concerns (e.g., carbon credit
trading/monetization).
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] FIG. 1 represents the effect of the gas reducing
oligosaccharide composition ("GROC") on total gas production during
in vitro incubation of ruminal contents supplemented with 0.02 g
ground alfalfa.
[0025] FIG. 2 represents the effect of GROC on hydrogen production
during in vitro incubation of ruminal contents supplemented with
0.02 g ground alfalfa.
[0026] FIG. 3 represents the effect of GROC on methane production
during in vitro incubation of ruminal contents supplemented with
0.02 g ground alfalfa.
[0027] FIG. 4 represents the effect of GROC on carbon dioxide
production during in vitro incubation of ruminal contents
supplemented with 0.02 g ground alfalfa.
[0028] FIG. 5 represents the effect of GROC on final pH during in
vitro incubation of ruminal contents supplemented with 0.02 g
ground alfalfa.
[0029] FIG. 6 represents the effect of GROC on ammonia accumulation
during in vitro incubation of ruminal contents supplemented with
0.02 g ground alfalfa.
DETAILED DESCRIPTION
[0030] Although an illustrative implementation of one or more
embodiments may be provided below, the disclosed systems and/or
methods may be implemented using any number of techniques. This
disclosure should in no way be limited to the illustrative
implementations, drawings, and techniques illustrated below,
including the exemplary designs and implementations illustrated and
described herein, but may be modified within the scope of the
appended claims along with their full scope of equivalents.
[0031] Disclosed herein is a gas reducing oligosaccharide
composition ("GROC") and methods of making and using same. In an
embodiment, the gas comprises any gas produced by and subsequently
expelled from an organism of the type to be described in more
detail later herein. Alternatively, the gas comprises methane,
ammonia, or combinations thereof. In an embodiment, the GROC
comprises soluble extractable materials from a lignocellulosic
source. In an embodiment, the GROC is derived from a renewable,
biological source material such as wood, bark, foliage, and roots.
As is understood by those of skill in the art, wood refers to the
organic material produced as secondary xylem in the stems of trees
comprising various biological polymers including cellulose,
hemicellulose, pectin, and lignin.
[0032] In an embodiment, the GROC is derived from a lignocellulosic
source material. Herein the term "derived" refers to isolation of
the material from an organism where it is present natively such
that the material is no longer in contact with all components of
its native milieu. In an embodiment, the GROC is derived from the
above-ground or below-ground portion of a plant source material. In
another embodiment the GROC is derived from a lignocellulosic
source material that exhibits cambial growth. For example, the
source material may comprise a plant that is a member of the order
Pinales, the family Pinaceae, the family Fagaceae or the order
Saxifragales. Alternatively the source material is derived from a
member of the family Pinaceae. The family Pinaceae comprises
coniferous trees commonly known as the pine family.
[0033] In embodiments, the source material comprises a material
derived from a member of the genus Pinus. The genus Pinus comprises
coniferous trees commonly known as the pines. In embodiments, the
source material comprises a material derived from a member of at
least one species collectively referred to as the Southern Yellow
Pines. In embodiments, the source material comprises a material
derived from a member of the species Pinus taeda L, and its hybrids
commonly referred to as Loblolly Pine. In alternative embodiments,
the source material comprises a material derived from a member of
the species Pinus elliotii Englem, and its hybrids commonly
referred to as the Slash Pine. In alternative embodiments, the
source material comprises a material derived from a member of the
species Pinus echinata Mill, and its hybrids commonly referred to
as Shortleaf Pine. In alternative embodiments, the source material
comprises a material derived from a member of the species Pinus
palustris Mill, and its hybrids commonly referred to as the
Longleaf Pines. Southern Yellow Pines of the type disclosed herein
are native to the Southeast United States and may typically be
found along the coastal plain from eastern Texas to southeast
Virginia extending into northern and central Florida. These
Southern Yellow Pines are also globally cultivated and as such it
is contemplated other regions may also provide a source of such
pines. Typically Southern Yellow Pines are characterized as having
a height of 30-35 m (100-115 ft) and a diameter of 0.7 m (28 in)
and may grow to 47 m (154 ft) with a diameter of 1.2 m (47 in).
Southern Yellow Pines may also be characterized by bark that is
thick, reddish-brown, and scaly and leaves that are dark green,
needle-like, and occur in bundles of up to three. The leaves are
often twisted and have a length ranging from 20-45 cm (8-18
in).
[0034] In embodiments, a process of deriving a GROC from a source
material (e.g., wood) comprises comminuting the source material,
extracting soluble material from the source material, and
concentrating the extracted solubles. In an embodiment, a process
of deriving a GROC from a source material (e.g., wood) comprises
comminuting the wood, extracting oligosaccharides and
polysaccharides (e.g., hemicellulose) from the source material via
contact with a solvent (e.g., water), and concentrating the solvent
extract.
[0035] In an embodiment, the process of deriving a GROC from a
source material optionally comprises comminution of the source
material to reduce the physical size of the source material. For
example, the wood source material may be chipped or comminuted
prior to extracting the soluble material. As will be appreciated by
those of skill in the art, comminuting the wood source material is
an appropriate means of reducing the wood to a size that is both
manageable and efficient for continued processing. Suitable
machinery known to those of skill in the art may be employed to
comminute the source material, non-limiting examples of which
include tub grinders, wood chippers, chip-n-saws and the like.
Further, the comminuted wood may be screened to ensure that the
material is uniformly or substantially uniformly sized. In the
following embodiments, it is presumed that the wood source material
has been comminuted prior to further processing. Though one or more
of the following embodiments may describe the performance of
processes with respect to comminuted wood, it is specifically
contemplated that comminution is not necessarily a prerequisite to
these processes.
[0036] In an embodiment, the process of deriving a GROC from a
source material (e.g., wood) comprises extracting the soluble
material from the wood. Any method known to one of ordinary skill
in the art and not deleterious to the GROC may be employed to
extract the soluble material from the wood. In an embodiment, the
process of extracting the soluble material from the wood comprises
softening the source material (e.g., wood), optionally comminuting
the softened wood, and contacting the softened wood with one or
more solvents into which the soluble material may partition. Herein
"softening" refers to processes which decrease the structural
integrity of the exposed cell walls of the source material.
[0037] In an embodiment, the source material (e.g., wood) is
softened using any methodology known to one of ordinary skill in
the art and compatible with the components of the GROC. Nonlimiting
examples of such methodologies include thermal, thermomechanical,
thermochemical, mechanical, chemical, hydrothermal, acid
hydrolysis, alkaline hydrolysis, organosolvent treatment, enzyme
treatment, or combinations thereof. In an embodiment, the
methodology comprises steam explosion and decompression wherein the
source material is subjected to steam, pressure, and elevated
temperature for some specified time period to soften and dissolve
cell wall constituents.
[0038] In an embodiment, the source material is softened by a
technique comprising autohydrolysis. As used herein, the term
"autohydrolysis" refers to the process of subjecting the source
material to a high temperature in the absence of chemicals but with
moisture wherein organic acids are formed from functional groups
such as acetyl groups liberated from the source material.
[0039] Specifically, the autohydrolysis process may comprise
introducing the source material (e.g. comminuted wood) into a steam
digester. In embodiments, the comminuted wood is steamed at a
pressure ranging from 18-300 psi, alternatively, from 50-250 psi,
alternatively, from 75-225 psi. In embodiments, the comminuted wood
will be allowed to remain in the steam digester for a period up to
10 minutes, alternatively, up to 15 minutes, alternatively, up to
20 minutes. In an embodiment, temperatures within the steam
digester range from 212-420.degree. F., alternatively, from
290-340.degree. F., alternatively, from 295-335.degree. F.,
alternatively, from 300-330.degree. F. Not seeking to be bound by
any particular theory, introduction into the steam digester softens
the woods chips, thereby increasing the efficiency of later
processing steps which seek to extract the soluble material.
[0040] In an embodiment, the source material is softened by a
technique comprising pulping. Any pulping process known to one of
ordinary skill in the art and not deleterious to the GROC may be
employed to soften the source material. Examples of such processes
are described in greater detail below.
[0041] In an embodiment, the source material (e.g., comminuted
wood) is pulped using a mechanical pulping process. In these
embodiments, the mechanical pulping process comprises separating
the component wood fibers via the use of a plurality of
grindstones, refining discs, knives, and like machinery known to
those of skill in the art to mechanically disintegrate the
comminuted wood, thereby reducing the comminuted wood to the
fibrous components.
[0042] In an embodiment, the source material is pulped by
subjecting the material to a pulping agent. In these embodiments,
the pulping process comprises subjecting the comminuted wood to one
or more chemicals and/or enzymes which will break down the lignin
that holds the fibrous components together. Thus, as the lignin is
degraded, the fibers of the wood are separated. Nonlimiting
examples of chemical pulping processes include acid hydrolysis,
alkaline hydrolysis, organosolvent treatment and the like.
[0043] In some embodiments other methodologies for softening the
source material may be employed. Such methodologies may employ a
variety of reaction parameters such as temperature, pressure, pH,
varying reaction times and the like to extract the soluble material
from the wood. For example, the source material may be softened by
a steam extrusion process. Herein steam extrusion refers to a
process wherein the source material (e.g., comminuted wood) is
pressed through a die where compressed gases (e.g., steam) are
developed and then expanded (released).
[0044] Hereinafter the source material whether subjected to a
process of the type described herein (e.g., optional comminution
followed by autohydrolysis or pulping) is termed the refined source
material and for simplicity will hereinafter be referred to as the
"refined wood."
[0045] In some embodiments, the process further comprises
comminuting the refined wood. Comminution and methods of carrying
out same have been described previously herein and may likewise be
used to reduce the size of the refined wood. The comminuted,
refined wood may be passed for washing as described below.
[0046] The process of deriving a GROC from a source material may
further comprise washing the refined wood. The refined wood may be
washed by contacting the material with a wash solution. The wash
solution may comprise any material compatible with the components
of the GROC. In an embodiment, the wash solution is an aqueous
solution; alternatively the wash solution is water or consists
essentially of water. Contacting of the refined wood and wash
solution may be carried out using any suitable technique such as
for example by showering the refined wood with a wash solution. As
the refined wood is contacted with the wash solution the
extractable compounds may be dissolved in or otherwise portioned
into the wash solution which may then be collected. In an
embodiment, the soluble material comprising oligosaccharides and
polysaccharides (e.g., hemicellulose) present in the refined wood
will be dissolved, suspended in, or otherwise partitioned into the
wash solution.
[0047] In some embodiments, softening of the source material and
extraction of the soluble material may be carried out concomitantly
using a process such as solid-liquid countercurrent extraction.
Herein, solid-liquid countercurrent extraction refers to a process
wherein a solid phase material (e.g., comminuted wood) and a liquid
phase material (e.g., hot water) are contacted to each other by
causing them to flow countercurrently to each other to adsorb part
of the components contained in the liquid phase to the solid phase
and simultaneously extract part of the components adsorbed to the
solid phase into the liquid phase.
[0048] The wash solution obtained by the processes described herein
comprises soluble material extractable from a source material of
the type described previously herein. Hereinafter the wash solution
obtained as described is termed the soluble extractable material
("SEM"). In an embodiment, processes of the type described herein
result in the extraction of greater than about 50% of the
hemicellulose present in the source material, alternatively greater
than about 60, 65, 70, 75, or 80% of the hemicellulose present in
the source material.
[0049] In an embodiment, the SEM may be further processed by
concentrating the solution to form a concentrated liquid. In
embodiments, the SEM is concentrated to between 40 and 70% solids,
alternatively to between 12% to 40% solids, alternatively to
between 70% to 90% solids. The solids found in the SEM comprise
approximately 93% carbohydrate material, approximately 4% ash, and
less than approximately 1% each of protein, fat, or crude fiber and
exhibit methane-reducing activity.
[0050] In an embodiment, the SEM is dehydrated to remove excess
moisture. The SEM may be dehydrated using any suitable dehydration
process as known to those of skill in the art and compatible with
the needs of the process (e.g., spray drying, drum drying). In an
embodiment, the SEM may be dehydrated to a moisture content of less
than about 18%, alternatively less than about 10%, alternatively
less than about 5%. In an embodiment, the SEM is concentrated
and/or dehydrated to yield a solids powder.
[0051] The SEM prepared as described herein may comprise
monosaccharides, oligosaccharides and polysaccharides. The term
oligosaccharide herein refers to a polymer comprising from about 2
to about 20 monosaccharide units while a polysaccharide herein
refers to a polymer comprising greater than about 20 monosaccharide
units. The number of monosaccharide units in a given
oligosaccharide is termed the "degree of polymerization" (DP). For
example, the SEM may comprise polysaccharides having a DP of
greater than about 100, alternatively greater than about 150, 200,
250, 300, 350, 400, 450, or 500. In an embodiment, the SEM may
comprise monomers, oligosaccharides, and polymers ranging from
about 2 to about 500 DP as will be described in more detail later
herein.
[0052] In embodiments the SEM comprises one or more
oligosaccharides comprising a polysaccharide backbone; that is, the
backbone comprises a plurality of glycosidically-linked
monosaccharide units. In embodiments, the glycosidic linkage
comprises an .alpha.-glycosidic link, a .beta.-glycosidic link, or
combinations thereof. In embodiments, the SEM comprises
oligosaccharides comprising both .alpha.-glycosidic links and
.beta.-glycosidic links. In embodiments, the oligosaccharide will
further comprise at least one side-chain. The side chain may
comprise at least one monosaccharide unit glycosidically-linked to
at least one saccharide unit of the polysaccharide backbone.
Alternatively, the side chain may comprise at least one
polysaccharide unit glycosidically-linked to at least one
saccharide unit of the polysaccharide backbone.
[0053] In embodiments, the SEM comprises one or more
oligosaccharides having monomeric units comprising an aldotriose
monomer, an aldotetrose monomer, an aldopentose monomer, an
aldohexose monomer, a ketotriose monomer, a ketotretrose monomer, a
ketopentose monomer, a ketohexose monomer, a ribose monomer, an
arabinose monomer, a xylose monomer, a lyxose monomer, an allose
monomer, an altrose monomer, a glucose monomer, a mannose monomer,
a gulose monomer, an idose monomer, a galactose monomer, a talose
monomer, a ribulose monomer, a xylulose monomer, a psicose monomer,
a fructose monomer, a sorbose monomer, a tagatose monomer, or
combinations thereof.
[0054] In an embodiment, the SEM is further processed to reduce the
DP of the constituent polymers. The DP of the SEM constituent
polymers (e.g., polysaccharides) may be reduced by cleaving one or
more of the glycosidic bonds between the monomer units of an
oligosaccharide. Various methods can be used to cleave some of the
glycosidic bonds between the monomer units while preserving the
integrity of the sugar units. For example, the glycosidic bonds may
be hydrolyzed. Hydrolysis of the glycosidic bonds can be achieved
through any mechanism known to one of ordinary skill in the art and
compatible with the needs of the process. For example hydrolysis of
the glycosidic bonds may be carried out employing chemical,
enzymatic, thermal, or ultrasonic processes. Process variables such
as reagent concentration, pH, temperature, time, and reactant can
determine the degree of hydrolysis. Thus, one of ordinary skill in
the art with the benefits of this disclosure may select hydrolysis
reaction conditions suitable for the production of specific polymer
chain lengths.
[0055] In embodiments, the DP of the SEM constituent polymers is
reduced by acid hydrolysis of the material. For example, an acid
for cleaving glycosidic bonds suitably comprises a weak acid.
Non-limiting examples of such a weak acid include trifluoroacetic
acid (TFA), acetic acid, and oxalic acid. Alternatively, in
embodiments, an acid for cleaving glycosidic bonds suitably
comprises a strong mineral acid. Non-limiting examples of such a
strong mineral acid include sulfuric acid and hydrochloric acid. In
various embodiments, numerous combinations of exposure time,
temperature, and acid concentration can be used to hydrolyze any
large DP hemicellulose polysaccharides to the DP ranges disclosed
herein.
[0056] In alternative embodiments, the DP of the SEM constituent
polymers is reduced enzymatically. For example, enzymes may be
employed to cleave the polymer chains at specific linkages.
Numerous enzymes, including but not limited to .beta.-mannanase and
glucosidases, are suitable for use. Such enzymes and reaction
conditions suitable for enzymatic cleavage of the SEM would be
known to one of ordinary skill in the art with the aid and benefits
of this disclosure.
[0057] Hydrolysis of the SEM as described herein produces a
material hereinafter termed the "hydrolyzed hemicellulose material"
("HHM"). The HHM may have a DP of about 2 to about 30,
alternatively about 2 to about 20, alternatively about 2 to about
15, alternatively about 2 to about 12. In an embodiment, the HHM
comprises oligosaccharides having from about 3 to about 5 DP,
alternatively from about 9 to about 14 DP, alternatively from about
16 to about 18 DP.
[0058] In an embodiment, the HHM or the SEM is further processed by
contacting the material with a precipitating agent. Upon contact
with a precipitating agent, HHM/SEM-derived oligosaccharide
fractions having gas-reducing functionality of the type described
herein may be precipitated from the solution. In embodiments, a
material containing gas-reducing activity is precipitated from the
HHM or SEM when the HHM or SEM is contacted with a precipitating
agent comprising an alcohol. Alternatively, a material containing
gas-reducing activity is precipitated from the HHM or SEM when the
HHM or SEM is contacted with ethanol. Further processing of the
mixture comprising the precipitant may include removing the
precipitating agent (e.g., ethanol) using any suitable technique
(e.g., evaporation). The resulting precipitated material,
hereinfter termed the precipitated gas-reducing material ("PGM"),
may be dried or re-suspended in an appropriate solvent.
[0059] Additional processing of the PGM may involve subjecting the
material to enrichment methods in order to concentrate fractions
having a specific DP or remove non-active (e.g.,
non-methane-reducing) compounds. In embodiments, the PGM is further
enriched by subjecting the previously described SEM and/or its
derivatives (e.g., HHM) to additional separation procedures. In
these embodiments, such separation procedures include but are not
limited to chromatographic separation, ion exchange separation,
filtration, microfiltration, ultra filtration, or the like. Such a
separation process may be employed to remove any remaining
non-desirable materials (e.g., monosaccharide, lignin, salts,
phenolics, ash, etc.) from the product composition. Additionally
compounds, such as phenolics or lignin, may be removed at various
points during processing.
[0060] In an embodiment, the SEM, HHM, and/or PGM comprise
hemicellulose comprising xylans, arabinoxylans,
galactoglucomannans, or combinations or derivatives thereof. In an
embodiment, the SEM, HHM, and/or PGM comprise xylans. In some
embodiments, the xylan is comprised of a backbone chain of xylose
units which are linked by .beta.-(1,4)-glycosideic bonds and
branched by .alpha.-(1,2)-glycosidic bonds with
4-O-methylglucoronic acid groups. In some embodiments, O-acetyl
groups replace the OH groups in the C2 and C3 groups. A partial
structure of a xylan is shown in Structure 1:
##STR00001##
[0061] In an embodiment the SEM, HHM, and/or PGM comprise an
arabinoxylan. Arabinoxylans consist of .alpha.-L-arabinofuranose
residues attached as branch-points to .beta.-(1.fwdarw.4)-linked
D-xylopyranose polymeric backbone chains. These may be C2 or
C3-substituted or C2 and C3-di-substituted. The arabinose residues
may also be linked to other groups attached such as glucuronic acid
residues, ferulic acid crosslinks and acetyl groups. The most
stable conformations comprise .alpha.-L-arabinofuranose and
.beta.-(1.fwdarw.4)-linked D-xylopyranose residues. The furanose
can, however, take up a number of other conformations with similar
energy whereas the chair conformation of the pyranose residue is
fixed. Arabinoxylans may comprise greater than about 500
monosaccharide repeating units, alternatively greater than about
1000 monosaccharide repeating units, alternatively from about 1500
to about 5000 monosaccharide repeating units. A partial structure
of an arabinoxylan is shown in Structure 2:
##STR00002##
[0062] In embodiments, the SEM, HHM, and/or PGM comprise an
oligosaccharide comprising monomeric units having glucose monomers,
galactose monomers, and mannose monomers in the form of a
galactoglucomannan ("GGM"). In embodiments, the GGM comprises a
backbone of .beta.-1-4 linked mannose units with randomly spaced
glucose units included and occasional .alpha.-1-6 galactose unit
side chains. In embodiments, the hydroxyl groups of one or more
monomeric units comprising the GGM backbone are partially
substituted with O-acetyl groups at C-2 and C-3 positions. A
non-limiting representative GGM structure is shown in Structure
3:
##STR00003##
[0063] In an embodiment, the GGM oligosaccharide comprises glucose,
galactose, and mannose in a ratio of 3 to 1 to 6 respectively.
[0064] As will be understood by one of ordinary skill in the art,
variations in the methodology for obtaining the SEM, HHM, and/or
PGM may result in variations in the amounts and/or nature of the
components of the SEM, HHM, and/or PGM. Hereinafter the GROC which
may comprise the SEM, HHM, and/or PGM may be administered to an
organism in order to reduce gas production of the organism. In an
embodiment, the organism has a gastrointenstinal tract. In some
embodiments, the organism is a ruminant animal. Herein a ruminant
animal refers a mammal of the order Artiodactyla that digests
plant-based food by initially softening it within the animal's
first stomach, known as the rumen, then regurgitating the
semi-digested mass, now known as cud, and chewing it again.
[0065] In practical use, a GROC can be combined as the active
ingredient in intimate admixture with a pharmaceutical carrier
according to conventional pharmaceutical compounding techniques.
The carrier may take a wide variety of forms depending on the form
of preparation desired for administration. In preparing the
compositions for oral dosage form, any of the usual pharmaceutical
media may be employed, such as, for example, water, glycols, oils,
alcohols, flavoring agents, preservatives, coloring agents and the
like in the case of oral liquid preparations, such as, for example,
suspensions, elixirs and solutions; or carriers such as starches,
sugars, microcrystalline cellulose, diluents, granulating agents,
lubricants, binders, disintegrating agents and the like in the case
of oral solid preparations such as, for example, powders, capsules
and tablets. Because of their ease of administration, tablets and
capsules represent the most advantageous oral dosage unit form in
which case solid pharmaceutical carriers may be employed. If
desired, tablets may be coated by standard aqueous or nonaqueous
techniques.
[0066] Pharmaceutical compositions comprising a GROC suitable for
oral administration may be presented as discrete units such as
capsules, cachets or tablets each containing a predetermined amount
of the active ingredient (e.g., GROC), as a powder or granules or
as a solution or a suspension in an aqueous liquid, a non-aqueous
liquid, an oil-in-water emulsion or a water-in-oil liquid emulsion.
Such compositions may be prepared by any of the methods of pharmacy
but all methods include the step of bringing into association the
active ingredient with the carrier which constitutes one or more
necessary ingredients. In general, the compositions are prepared by
uniformly and intimately admixing the active ingredient with liquid
carriers or finely divided solid carriers or both, and then, if
necessary, shaping the product into the desired presentation. For
example, a tablet may be prepared by compression or molding,
optionally with one or more accessory ingredients. Compressed
tablets may be prepared by compressing in a suitable machine, the
active ingredient in a free-flowing form such as powder or
granules, optionally mixed with a binder, lubricant, inert diluent,
surface active or dispersing agent. Molded tablets may be made by
molding in a suitable machine, a mixture of the powdered compound
moistened with an inert liquid diluent.
[0067] The GROC may be used in combination with other compositions
that are used in the treatment/prevention/suppression or
amelioration of the adverse health events for which a GROC of the
type described herein are useful.
[0068] In an embodiment, the GROC is administered to an organism of
the type previously described herein. Administration of the GROC
may comprise preparing the GROC in a suitable orally ingestible
form and providing the suitable orally ingestible form to the
organism. Suitable orally ingestible forms are discussed herein in
further detail, although other suitable ingestible forms and
methods of formulating same will be appreciable by those of skill
in the art with the aid of this disclosure.
[0069] In an embodiment, a suitable orally ingestible form
comprises a GROC incorporated within a food, feed, or fodder
product. The GROC may be incorporated within the food, feed, or
fodder product as a dry powder or a liquid. Non-limiting examples
of food, feed, or fodder products into which the GROC may be
incorporated include compound feeds and premixes such as pellets,
liquid feed, nuts, nuggets, oil cakes, press cakes, various meals
(e.g., fishmeal), or combinations thereof. Such food, feed, or
fodder product may be prepared by admixing or blending the GROC
with a suitable carrier or diluent. Non-limiting examples of
suitable carriers may include grass and other forage plants, plant
oils, seeds, grains, crop residues, sprouted grains, legumes,
alfalfa meal, soybean meal, cottonseed oil meal, linseed oil meal,
sodium chloride, cornmeal, molasses, urea, corncob meal, rice
kernel, and the like. The carrier promotes a uniform distribution
of the active ingredients in the finished feed into which the
carrier is blended. It thus may ensure proper distribution of the
active ingredient throughout the food, feed, or fodder product.
[0070] In an embodiment, a suitable orally ingestible form
comprises a GROC prepared as a nutritional supplement. Such a
nutritional supplement may be ingestible by an organism alone or
with another food, feed, fodder, forage product, snack, treat, or
enjoyment product. In various embodiments, nutritional supplements
may be prepared in a wet, semi-wet, or dry form. Nonlimiting
examples of suitable nutritional supplement forms include powders,
granules, syrups, and pills; other suitable forms will be known to
those of skill in the art with the aid of this disclosure. In an
embodiment, a nutritional supplement may be added to another food,
feed, fodder, or forage product. For example the nutritional
supplement may comprise a powder or syrup which is dispensed with
(e.g., poured onto) hay, pellets, forage, or the like.
Alternatively, in an embodiment a nutritional supplement is
provided without any other food or nutrient. For example, the
nutritional supplement may comprise a syrup, gel, block, or tub
which may be licked by an organism (e.g., from a tub or other
suitable dispenser) or water-soluble powder dissolved in water
provided for ingestion by the organism. Other suitable means of
dispensing a nutritional supplement will be appreciated by those of
skill in the art viewing this disclosure.
[0071] As will be appreciated by those of skill in the art, the
ingestible forms may be formulated for ingestion by one or more
organisms, non-limiting examples of which include livestock such as
cattle, swine, horses, sheep, goats, poultry, fish, domesticated
companionship species such as dogs, cats, fish, and rodents or
undomesticated wildlife such as deer, moose, elk, migratory and
non-migratory fowl, decapods, and fish.
[0072] In an embodiment, administration of a GROC improves the
overall health of the organism to which it is administered. In some
embodiments, the overall improved health of the organism may be
evidenced by an increase in biological functions such as nutrient
uptake, muscle growth, muscle development, weight gain, coat
growth, survival, or combinations thereof. In another embodiment
administration of the GROC to an organism results in an increased
yield in an organism derived commodity such as eggs, meat, milk,
wool, or combinations thereof.
[0073] In an embodiment, a ruminant animal when administered a GROC
of the type described herein may display increased ruminal
fermentation rates, increased dry matter digestion and a reduction
in methane production when compared to an otherwise similar
ruminant animal not administered a GROC. Without wishing to be
limited by theory, administration of GROC of the type described
herein to a ruminant animal may alter the pH of the ruminal
environment resulting from an increased production of volatile
fatty acids. In an embodiment, administration of GROC may
selectively stimulate and/or inhibit the activity of certain
microorganisms in the rumen. In an embodiment, administration of
GROC of the type described herein to a ruminant animal results in a
change in the production of specific volatile fatty acids, and in
the relative proportion of specific volatile fatty acids. A
decrease in the acetate:propionate ratio is consistent with reduced
methane production. In an embodiment, administration of GROC of the
type described herein to a ruminant animal results in a change in
hexose and/or pentose fermentation in the ruminant animal.
EXAMPLES
Example 1
[0074] In this example, the effects of a GROC of the type described
herein on microbial efficiency and metabolism were evaluated in
continuous culture rumen fermentation. Rumen inoculum was fermented
for ten days and samples were collected on days 8, 9, and 10. Three
replications were conducted per treatment. Fermentation parameters
were analyzed. The amount of propionic acid increased while the
amount of acetic acid decreased with the inclusion of 1% GROC over
the control. This was seen in the molar percentage as well and the
mmoles per day measurements. See TABLE 1 for data. These
fermentation parameters are consistent with the conditions
favorable to the reduction of methane in the rumen.
TABLE-US-00001 TABLE 1 Item Control With 1% GROC Dry Matter
Digestion (%) 61.8 68.6 Molar % Acetic Acid 63.2 60.0 Molar %
Propionic Acid 17.6 20.4 Acetic:propionic ratio 3.63 2.98 Acetic
acid (mmoles/day) 251 240 Propionic acid (mmoles/day) 70 82
Example 2
[0075] Freshly collected ruminal contents containing mixed
populations of ruminal bacteria were inoculated (1 g per tube) into
18.times.150 mm crimp top Hungate tubes filled with 9 ml anaerobic
basal broth. The basal medium contained essential minerals,
nutrients and vitamins was supplemented with finely ground alfalfa
(2.0% wt/vol) and buffered to pH 6.80. GROC was included in sets of
triplicate incubation doses at 0, 0.03, 0.10, 0.30 and 0.60 per 10
ml.
[0076] All incubations were conducted at 39.degree. C. under a 100%
CO.sub.2 gas phase for 24 h. After 24 h incubation, gas volumes
were measured by recording displacement of volume in a lubricated
glass syringe and 1 ml headspace gas samples were injected into a
gas chromatograph for determination of hydrogen, methane and carbon
dioxide composition (Allison et al., 1992). Aliquots from each
incubation tube were also measured for pH and for colorimetric
determination of ammonia concentrations (Chaney and Marbach, 1962).
Fluid samples collected at 0 and 24 h incubation were frozen and
shipped to the National Animal Disease Center in Ames, Iowa for
determination of volatile fatty acid accumulations by gas
chromatography (Salanitro and Muirhead, 1975). A subsequent study
was conducted similarly except using 0.02% (wt/vol) trypticase as
an added protein substrate to assess the potential impact of GROC
on protein and amino acid metabolism.
[0077] A general analysis of variance revealed main effects
(P<0.05) of GROC on final pH and on total volume and composition
of gas produced during in vitro incubation of mixed populations of
ruminal microbes (FIGS. 1-5). Quadratic trends were observed for
effects of GROC on accumulation of hydrogen and carbon dioxide with
the highest amounts of these gases being produced in incubations
supplemented with 0.10 g inclusion levels but with production
declining rapidly in incubations supplemented with >0.3 g
product. Linear effects of GROC were observed on pH and methane
production. These results suggest that the lower inclusion levels
of GROC had no direct negative effect on ruminal fermentation but
at the higher inclusion levels an indirect effect of the lower pH
likely inhibited gas production. This conclusion is supported by
the observation of increasing amounts of formate, acetate, lactate
and succinate produced in incubations supplemented with greater
amounts of GROC. See TABLE 2. The production of these volatile
fatty acids typically increases with readily fermentable
substrates. The quadratic responses observed with respect to the
production of the more reduced volatile fatty acids propionate,
butyrate and valerate are not unexpected as these acids are
inversely correlated with lactate production. Similarly, formate
and succinate generally do not accumulate in ruminal fermentations
unless methane production is decreased. A linear increase in
amounts of hexose fermented was observed with increasing inclusion
of GROC which indicates the product contained appreciable
quantities of readily fermentable carbohydrate.
[0078] Using 0.02% tryptose, an enzymatic digest of soybean meal,
to assess the potential impact of GROC on protein and amino acid
metabolism has been analyzed. Results reveal that rates of ammonia
accumulation (not shown) and the total amount of ammonia produced
during incubation with added tryptose decreased with increasing
GROC supplementation (FIG. 6). Rates of ammonia production by
ruminal microbes are markedly influenced by pH with rates being
highest near pH 8.0 and declining rapidly at pH<7.0. The pH
measured at the end of the tryptose incubations declined linearly
(P<0.05) with increasing GROC supplementation (6.61.+-.0.02,
6.45.+-.0.03, 5.98.+-.0.05, 4.66.+-.0.04 and 4.32.+-.0.01 for
incubations containing 0, 0.03, 0.10, 0.30 and 0.60 g added
product, respectively) indicating that pH may have influenced
protein catabolism in these incubations.
TABLE-US-00002 TABLE 2 Volatile fatty acid production and
stoichiometric estimation of hexose fermentation during in vitro
incubation of increasing concentrations of GROC with mixed
populations of ruminal microbes in freshly collected rumen fluid
supplemented with 0.2 g ground alfalfa. Treatment Volatile fatty
acid production.dagger. None 0.03 0.10 0.30 0.60 SEM P Formate
(.mu.mol ml.sup.-1) <0.15.sup.c <0.15.sup.c 1.45.sup.c
10.77.sup.b 19.10.sup.a 0.58 <0.0001 Acetate (.mu.mol ml.sup.-1)
42.57.sup.b 40.79.sup.b 53.53.sup.b 56.87.sup.ab 73.96.sup.a 4.00
0.0011 Propionate (.mu.mol ml.sup.-1) 22.52.sup.c 31.59.sup.b
54.15.sup.a 24.22.sup.c 6.37.sup.d 1.24 <0.0001 Butyrate
(.mu.mol ml.sup.-1) 3.74.sup.b 3.83.sup.b 9.88.sup.a 1.47.sup.b
0.31.sup.b 0.84 0.0001 Lactate (.mu.mol ml.sup.-1) <0.15.sup.c
<0.15.sup.c <0.15.sup.c 76.32.sup.b 109.31.sup.a 0.20
<0.0001 Valerate (.mu.mol ml.sup.-1) 0.51.sup.b 0.24.sup.b
3.01.sup.a 0.17.sup.b <0.15.sup.b 0.14 <0.0001 Isobutyrate
(.mu.mol ml.sup.-1) 0.21.sup.a 0.15.sup.b <0.15.sup.b
<0.15.sup.b <0.15.sup.b 0.37 0.0050 Isovalerate (.mu.mol
ml.sup.-1) <0.15.sup.b <0.15.sup.b <0.15.sup.b
<0.15.sup.b 0.21.sup.a 0.02 <0.0002 Succinate (.mu.mol
ml.sup.-1) 0.96.sup.c 0.64.sup.c 3.14.sup.bc 9.91.sup.a 4.74.sup.b
0.63 <0.0001 Total (.mu.mol ml.sup.-1) 70.67.sup.d 77.33.sup.d
125.42.sup.c 180.40.sup.b 215.01.sup.a 5.95 <0.0001
Acetate:Propionate ratio 1.89.sup.bc 1.29.sup.bc 0.99.sup.c
2.35.sup.b 11.60.sup.a 0.25 <0.0001 Stoichiometric
calculations.sup..dagger-dbl. Hexose fermented (.mu.mol ml.sup.-1)
15.91.sup.d 26.07.sup.c 49.53.sup.ab 43.89.sup.b 53.63.sup.a 1.81
<0.0001 .dagger.Tests for effect of GROC level were conducted
via a general analysis of variance. Values are the mean from
cultures incubations in triplicate. .sup..dagger-dbl.Amounts of
hexose fermented were calculated as 1/2 acetate + 1/2propionate +
butyrate + valerate (DeMeyer, 1991). .sup.a,b,c,dMeans within rows
with unlike superscripts differ (P < 0.05) based on a Tukeys
All-Pairwise Comparison test.
[0079] As can be seen from Example 2, the amount of methane
produced significantly decreased as the amount of composition
increased.
[0080] To determine a gas reducing composition dosing regime, a
dosing factor may be utilized. Dosing factors may be calculated
utilizing accepted/customary methodologies and/or procedures to
derive the desired and/or effective amount of GROC to be provided
to a subject animal.
[0081] The dosing factor may be in the range of about 0.0001 g/ml
to 0.1000 g/ml. In embodiments the dosing factor may be about
0.0001 g/ml, 0.0002 g/ml, 0.0005 g/ml, 0.0010 g/ml, 0.0015 g/ml,
0.0020 g/ml, 0.0025 g/ml, 0.0030 g/ml, 0.0035 g/ml, 0.0040 g/ml,
0.0045 g/ml, 0.0050 g/ml, 0.0055 g/ml, 0.0060 g/ml, 0.0065 g/ml,
0.0070 g/ml, 0.0075 g/ml, 0.0080 g/ml, 0.0085 g/ml, 0.0090 g/ml, or
0.0095 g/ml. In other embodiments, the dosing factor may be about
0.0100 g/ml, 0.0150 g/ml, 0.0200 g/ml, 0.0250 g/ml, 0.0300 g/ml,
0.0350 g/ml, 0.0400 g/ml, 0.0450 g/ml, 0.0500 g/ml, 0.0550 g/ml,
0.0600 g/ml, 0.0650 g/ml, 0.0700 g/ml, 0.0750 g/ml, 0.0800 g/ml,
0.0850 g/ml, 0.0900 g/ml, 0.0950 g/ml, or 0.1000 g/ml. In all of
the above-stated embodiments, the variance in the stated values may
range from about 1% to 50%.
[0082] In an embodiment, test doses of 0.03 g, 0.10 g, 0.30 g, and
0.60 g of GROC per 10 ml incubation fluid correspond to GROC dosing
factors of 0.003 g/ml, 0.01 g/ml, 0.03 g/ml, and 0.06 g/ml,
respectively. As such, to determine the appropriate dosing regime,
the volume of the embodiment's subject ruminant's rumen (or other
subject animal's digestive system contents) may be multiplied by a
representative GROC dosing factor of 0.003 g/ml, 0.01 g/ml, 0.03
g/ml, or 0.06 g/ml to correlate the dosage amounts of the GROC
tested in vitro to in vivo amounts. That resulting value may then
be divided by the mass of the subject to determine an amount of
GROC per unit of body weight.
[0083] As will be understood by one of ordinary skill in the art,
the amount of a composition (e.g., GROC) utilized to observe an in
vitro response may differ significantly from that required to
observe an in vivo response of the same type and magnitude.
Particularly, the determination of in vivo dosing amounts and
regimes is a multifactorial analysis that may be undertaken by the
ordinarily skilled artisan using any suitable methodology. This
disclosure contemplates determination of in vivo dosing amounts and
regimens effective to produce the beneficial properties disclosed
herein (e.g., reduction in methane production by an organism).
Further this disclosure contemplates the in vivo dosing amounts
effective to produce the beneficial properties disclosed herein may
differ significantly from the in vitro dosing amounts disclosed to
produce beneficial properties of the same type and magnitude.
[0084] For example, assuming a mature ruminant has a mass of
approximately 500 kg and has a rumen volume of 60 liters (60,000
ml), the in vitro test concentrations are multiplied by a 60,000 ml
rumen volume to yield 180 g, 600 g, 1800 g, and 3600 g per
ruminant, respectively. Thus, for a ruminant having a mass of 500
kg, the corresponding doses would be administered as 0.36 g/kg, 1.2
g/kg, 3.6 g/kg, and 7.2 g/kg, respectively, to correlate the dosage
concentrations of the GROC tested in vitro to in vivo amounts.
[0085] Alternatively, the percentage of dry matter consumed
relative to body mass may be utilized to calculate the amount of
GROC to be added to an amount of dry matter to achieve the desired
GHG production reduction. In an embodiment, a ruminant may consume
2.5% of its body mass per day in dry matter (e.g., feed or other
dietary intake). Thus, to determine the appropriate GROC dosing
regime, the subject animal's mass and rumen volume may be used in
conjunction with the subject animal's daily dry matter intake to
calculate the amount of GROC to be added to an amount of dry matter
to achieve the desired GHG production reduction.
[0086] For example, considering a hypothetical ruminant having a
mass of 500 kg and a rumen volume of 60 liters, representative GROC
dosing factors of 0.003 g/ml, 0.01 g/ml, 0.03 g/ml, or 0.06 g/ml
would correspond to in vivo GROC amounts of 180 g, 600 g, 1800 g,
and 3600 g, respectively. As such, again considering that a
ruminant consumes 2.5% of its body mass per day in dry matter, the
corresponding amount consumed for the 500 kg ruminant would be
12,500 g of dry matter. Thus, calculating the percentage of GROC
included in the ruminant's diet corresponding to the dosage
concentrations of the GROC tested in vitro, the corresponding
calculated in vivo amounts of GROC (e.g., 180 g, 600 g, 1800 g, and
3600 g) should be divided by the calculated amount of dry matter
(e.g., 12,500 g) to derive the appropriate percentage of GROC to be
supplemented by mass to the ruminant's diet.
[0087] Accordingly, the percent supplementation of GROC for the
above hypothetical ruminant would be: 180 g GROC/12500 dry
matter=1.4% GROC supplementation; 600 g GROC/12500 g dry
matter=4.8% GROC supplementation; 1800 g GROC/12500 g dry
matter=14.4% GROC supplementation; and 3600 g product/12500 g dry
matter=28.8% GROC supplementation. These percentages, 1.4%, 4.8%,
14.4%, and 28.8% for in vivo dry matter supplementation correspond
to the achieved percentage gas production effects indicated in
FIGS. 1-4 for the in vitro test amounts of GROC 0.03 g, 0.10 g,
0.30 g, and 0.60 g, respectively.
[0088] Specifically, for an embodiment, a 14.4% supplementation of
GROC in a ruminant's dry matter diet would result in about a 12%
decrease in the amount of total ruminal gas produced by said
ruminant. A 28.8% supplementation of GROC in a ruminant's dry
matter diet would result in about a 35% decrease in the amount of
total ruminal gas produced by said ruminant. These reductions are
surprising and unexpected because both a 1.4% and 4.8%
supplementation of GROC in a ruminant's dry matter diet result in
increases in total ruminal gas production.
[0089] Specifically, for an embodiment, a 1.4% supplementation of
GROC in a ruminant's dry matter diet would result in about a 30%
decrease in the amount of ruminal methane produced by said
ruminant. A 4.8% supplementation of GROC in a ruminant's dry matter
diet would result in about a 50% decrease in the amount of ruminal
methane produced by said ruminant. A 14.4% supplementation of GROC
in a ruminant's dry matter diet would result in a 95% decrease in
the amount of ruminal methane produced by said ruminant. A 28.8%
supplementation of GROC in a ruminant's dry matter diet would
result in a 99% decrease in the amount of ruminal methane produced
by said ruminant. These reductions are surprising and unexpected
because both a 1.4% and 4.8% supplementation of GROC in a
ruminant's dry matter diet result in increases in total ruminal gas
production as well as carbon dioxide production while resulting in
simultaneous decreases in ruminal methane production.
[0090] Specifically, for an embodiment, a 14.4% supplementation of
GROC in a ruminant's dry matter diet would result in about a 12%
decrease in the amount of ruminal carbon dioxide produced by said
ruminant. A 28.8% supplementation of GROC in a ruminant's dry
matter diet would result in about a 66% decrease in the amount of
ruminal carbon dioxide produced by said ruminant. These reductions
are surprising and unexpected because both a 1.4% and 4.8%
supplementation of GROC in a ruminant's dry matter diet result in
increases in ruminal carbon dioxide production.
[0091] A supplementation/dosing regime could comprise supplementing
a ruminant's dry matter diet with up to about 50% of GROC to result
in each of a total ruminal gas production reduction, a ruminal
methane production reduction, and a ruminal carbon dioxide
production reduction.
[0092] A supplementation/dosing regime could comprise supplementing
a ruminant's dry matter diet with approximately 10% to 30% of GROC
to result in each of a total ruminal gas production reduction, a
ruminal methane production reduction, and a ruminal carbon dioxide
production reduction.
Example 3
[0093] In this example, the effects of a GROC of the type described
herein on fiber degradation rates were evaluated in 12 multiparous
Holstein cows (142.+-.44 days in milk, 685.+-.19 kg body weight)
including four with ruminal fistula were used in a 2.times.2 Latin
square with 21-d periods. Two diets were fed--(i) a control typical
Midwest diet containing 55:45 forage (2/3 corn silage, 1/3 alfalfa
hay) to concentrate ratio; and (ii) a treatment diet in which 1.0%
of the diet dry matter ("DM") was replaced with GROC. DM intake
averaged 27.1 and 26.9 kg/d for the control and treatment,
respectively, and was not affected by treatment.
[0094] In situ testing was performed using Dacron bags containing
corn silage, alfalfa hay, or control or treatment total mixed
ration ("TMR"). The bags were inserted in triplicate into the
rumens of the 4 fistulated cows, TMR corresponding to the current
diet. The bags were incubated for from 0 to 48 hours, and
degradation of forages and TMR were analyzed. The in situ fiber
disappearance data is shown in TABLE 3. The increase in fiber
degradation rates of forages and diets with the inclusion of GROC
demonstrates the ability of the material in affect ruminal
digestion and/or fermentation.
[0095] For corn silage, the rate of disappearance (Kd) of neutral
detergent fiber ("NDF") (1.7 vs. 4.3) and acid detergent fiber
("ADF") (1.8 vs. 4.7%/h) increased (P<0.05) for cows fed the
treatment diet.
[0096] For alfalfa hay, the disappearance of fraction A of DM, NDF,
and ADF decreased and fraction B of DM and NDF increased with
treatment (P<0.05). The Kd for DM (8.0 vs. 11.0), NDF (6.3 vs.
10.3), and ADF (5.5 vs. 9.2) increased greatly for the alfalfa hay
in rumens of treated cows (P<0.05).
[0097] The results of EXAMPLE 3 demonstrate that supplementing
diets of lactating dairy cows with GROC has a beneficial effect on
fiber degradation characteristics.
TABLE-US-00003 TABLE 3 Control Treatment p< In Situ DM
Disappearance Alfalfa Hay A 32.4 26.7 0.04 B 35.2 41.6 0.06 Kd 8 11
0.05 In Situ NDF Disappearance Alfalfa Hay A 9.4 3.6 0.02 B 33.1
38.9 0.06 Kd 6.3 10.3 0.01 Corn Silage Kd 1.7 4.27 0.05 TMR A 9.6
20.2 0.01 In Situ ADF Disappearance Alfalfa Hay Kd 5.5 9.2 0.01
Corn Silage Kd 1.8 4.7 0.01 TMR A 11.4 17.1 0.03
[0098] An animal, when administered GROC, may display a reduction
in GHG production, e.g., a reduction in methane production and/or
carbon dioxide production, as compared to an otherwise similar or
same animal not administered GROC. The GHG reduction may be
associated with digestive activities of the animal, e.g., cud
formation, sustenance breakdown, manure deposition, or combinations
thereof.
[0099] As is evident in FIGS. 3 and 4, significant reductions in
the amounts of methane and carbon dioxide can be achieved via the
administration of GROC.
[0100] As shown in FIG. 3, the amount of ruminal methane produced
can be significantly reduced via the introduction of GROC into a
ruminant's diet. For example, FIG. 3 indicates that, depending on
the proportion of GROC administered in relation to the remainder of
the ruminant's diet, ruminal methane production can be decreased.
The decrease in ruminal methane production can be in the range from
about 1% to about 99%. For example, the decrease in ruminal methane
production can be from about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,
45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, up to about
99%. Preferably, the decrease in ruminal methane production can be
in the range from about 30% to about 99%.
[0101] As shown in FIG. 4, the amount of ruminal carbon dioxide
produced can be significantly reduced via the introduction of GROC
into a ruminant's diet. For example, FIG. 4 indicates that,
depending on the proportion of GROC administered in relation to the
remainder of the ruminant's diet, ruminal carbon dioxide production
can be decreased. The decrease in ruminal carbon dioxide production
can be in the range from about 1% to about 99%. For example, the
decrease in ruminal carbon dioxide production can be from about 5%,
10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,
75%, 80%, 85%, 90%, 95%, up to about 99%. Preferably, the decrease
in ruminal carbon dioxide production can be in the range from about
10% to about 66%.
[0102] The above reductions in carbon dioxide and methane may be
utilized by farmers and producers as resulting carbon credits.
Carbon credit as used herein is a generic term meaning that a value
has been assigned to a reduction or offset of GHG emissions for
sale, trading, or regulatory permitting and/or compliance
purposes.
[0103] Every GHG has a global warming potential ("GWP"), a
measurement of the impact that particular gas has on "radiative
forcing"; that is, the additional heat/energy which is retained in
the Earth's ecosystem through the addition of this gas to the
atmosphere.
[0104] The GWP of a given gas describes its effect on climate
change relative to a similar amount of carbon dioxide. As the base
unit, carbon dioxide's GWP numeric is 1.0. This allows regulated
GHGs to be converted to the common unit of carbon dioxide
equivalents ("CO.sub.2e"). For example, methane, a CO.sub.2e, has a
GWP of 21-meaning that one ton of methane will have an effect on
global warming that is 21 times greater than one ton of carbon
dioxide.
[0105] Carbon trading is an application of an emissions trading
approach. GHG emissions are capped and then markets are used to
allocate the emissions among the group of regulated sources. The
goal is to allow market mechanisms to drive industrial and
commercial processes in the direction of low emissions or less
carbon intensive approaches than those used when there is no cost
to emitting carbon dioxide and other GHGs into the atmosphere.
Since GHG reduction projects generate credits, this approach can be
used to finance carbon reduction schemes between trading partners
and around the world.
[0106] Climate exchanges have been established to provide a spot
market in allowances, as well as futures and options market to help
discover a market price and maintain liquidity. Currently there are
five exchanges trading in carbon allowances: the Chicago Climate
Exchange, European Climate Exchange, Nord Pool, PowerNext and the
European Energy Exchange.
[0107] Carbon prices are normally quoted in Euros per ton of carbon
dioxide or its carbon dioxide equivalent (CO.sub.2e). Other GHGs,
e.g., methane, can also be traded, but as indicated, are quoted as
standard multiples of carbon dioxide with respect to their GWP.
These features reduce a GHG's cap's financial impact on business,
while ensuring that the GHG's limits are met at a national and
international level.
[0108] Farmers and producers who supplement their livestocks' diets
with GROC may benefit from both international and national
emissions trading mechanisms by converting and/or applying their
livestocks' reductions in GHG, e.g., carbon dioxide and methane, to
carbon credits and then monetizing those carbon credits on the
appropriate climate exchanges.
[0109] For example, the United States Environmental Protection
Agency reports that globally, ruminant livestock produce about 80
million metric tons of methane annually, accounting for about 28%
of global methane emissions from human-related activities. In the
U.S., cattle emit about 5.5 million metric tons of methane per year
into the atmosphere, accounting for 20% of U.S. methane
emissions.
[0110] A single adult cow, by itself, may emit 80-110 kg of methane
per year. This means a farmer or producer with only 10 head of
cattle could be responsible for 1 metric ton of methane per year.
Accordingly, a farmer or producer with 100,000 head of cattle could
be responsible for 10,000 metric tons of methane per year. Thus, if
that farmer or producer were to utilize GROC in their livestock's
diet, that farmer or producer could reduce their livestock's
methane production by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,
50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, up to about
99%--i.e, tons of GHGs could be effectively prevented from entering
the environment/atmosphere.
[0111] Similarly, if that farmer or producer were to utilize GROC
in their livestock's diet, that farmer or producer could also
reduce their livestock's ruminal carbon dioxide production by 5%,
10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,
75%, 80%, 85%, 90%, 95%, up to about 99%--again, effectively
preventing tons of GHGs from entering the
environment/atmosphere.
[0112] Converting such percentage reductions into methane and/or
carbon dioxide tonnage reductions would allow said farmer or
producer to convert such reductions into carbon credits. In order
to convert the GHG production reductions into carbon credits, the
farmer or producer must first establish a baseline value for their
GHG producing activities. This baseline value would be determined:
(i) based on the amount of GHC emissions expected pursuant to
established emissions values and/or metrics for similarly situated
activities sans any GHG emissions reducing endeavors; (ii) based on
actually measured GHG emissions via accepted and/or established GHG
emission measuring protocols and procedures (prior to GHG reducing
endeavors); or (iii) by way of any other acceptable baseline
establishing method and/or procedure. GHG emission reductions could
then be quantified: (i) by projecting and/or extrapolating measured
and/or calculated in vitro or simulated GHG emission reducing
effects as in vivo/onsite reductions of GHG emissions; (ii) by
actually measuring GHG emissions via accepted and/or established
GHG emission measuring protocols and procedures; or (iii) by way of
any other acceptable quantification method and/or procedure.
[0113] A GHG emitting entity/facility may be subject to certain
designated/allowed levels of emissions for various types of GHG as
established or promulgated by an authorized emissions
governing/enforcing entity. As such, should said GHG emitting
entity/facility emit GHG at levels below the designated/allowed
levels, said GHG emitting entity/facility could convert its
designated/allowed yet non-emitted amounts of GHG to carbon
credits. For conversion purposes, one carbon credit is typically
considered equivalent to one metric ton of CO.sub.2 (or CO.sub.2e)
emissions.
[0114] The resulting carbon credits could then be traded on various
climate exchanges to effectively monetize the GROC's effect on
livestock for the economic/revenue benefit of the farmer or
producer.
[0115] Such monetization, coupled with GROC's abitlity to reduce
livestock-derived GHG while maintaining the livestock's feed intake
and protein synthesis, e.g., providing for optimal livestock
health, meat, and dairy production--unlike other similarly directed
plant-based feed additives, would allow farmers and producers to
maximize the economic output of their livestock management
operations.
[0116] For example, as a result of supplementing their livestock's
diets with GROC, farmers and producers would be able to optimize
the health, size, and output of their livestock (to maximize the
economic returns on such livestock's typical commodity-type
concerns) while also converting the GROC-derived reductions in GHGs
into carbon credits (to create and maximize an alternative
livestock economic/revenue concern).
[0117] Such methods and systems of realizing the synergistic
effects of GROC-supplemented livestock management benefits farmers
and producers, consumers of said livestock's products, and all
other persons concerned with reducing GHG emissions and protecting
the environment/atmosphere.
[0118] While embodiments of the invention have been shown and
described, modifications thereof can be made by one skilled in the
art without departing from the spirit and teachings of the
invention. The embodiments described herein are exemplary only, and
are not intended to be limiting. Many variations and modifications
of the invention disclosed herein are possible and are within the
scope of the invention. Where numerical ranges or limitations are
expressly stated, such express ranges or limitations should be
understood to include iterative ranges or limitations of like
magnitude falling within the expressly stated ranges or limitations
(e.g., from about 1 to about 10 includes, 2, 3, 4, etc.; greater
than 0.10 includes 0.11, 0.12, 0.13, etc.). For example, whenever a
numerical range with a lower limit, R.sub.L, and an upper limit,
R.sub.U, is disclosed, any number falling within the range is
specifically disclosed. In particular, the following numbers within
the range are specifically disclosed:
R=R.sub.L+k*(R.sub.U-R.sub.L), wherein k is a variable ranging from
1 percent to 100 percent with a 1 percent increment, i.e., k is 1
percent, 2 percent, 3 percent, 4 percent, 5 percent, . . . 50
percent, 51 percent, 52 percent, . . . , 95 percent, 96 percent, 97
percent, 98 percent, 99 percent, or 100 percent. Moreover, any
numerical range defined by two R numbers as defined in the above is
also specifically disclosed. Use of the term "optionally" with
respect to any element of a claim is intended to mean that the
subject element is required, or alternatively, is not required.
Both alternatives are intended to be within the scope of the claim.
Use of broader terms such as comprises, includes, having, etc.
should be understood to provide support for narrower terms such as
consisting of, consisting essentially of, comprised substantially
of, etc.
[0119] Accordingly, the scope of protection is not limited by the
description set out above but is only limited by the claims which
follow, that scope including all equivalents of the subject matter
of the claims. Each and every claim is incorporated into the
specification as an embodiment of the present invention. Thus, the
claims are a further description and are an addition to the
embodiments of the present invention. The discussion of any
reference herein is not an admission that it is prior art to the
presently disclosed subject matter, especially any reference that
may have a publication date after the priority date of this
application. The disclosures of all patents, patent applications,
and publications cited herein are hereby incorporated by reference,
to the extent that they provide exemplary, procedural or other
details supplementary to those set forth herein.
* * * * *