CA2155908C - Method for improving utilisation of nutrients by ruminant or ruminant-like animals - Google Patents
Method for improving utilisation of nutrients by ruminant or ruminant-like animals Download PDFInfo
- Publication number
- CA2155908C CA2155908C CA 2155908 CA2155908A CA2155908C CA 2155908 C CA2155908 C CA 2155908C CA 2155908 CA2155908 CA 2155908 CA 2155908 A CA2155908 A CA 2155908A CA 2155908 C CA2155908 C CA 2155908C
- Authority
- CA
- Canada
- Prior art keywords
- ruminant
- preparation
- animal
- whole cell
- group
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0001—Archaeal antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55566—Emulsions, e.g. Freund's adjuvant, MF59
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55583—Polysaccharides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Fodder In General (AREA)
Abstract
The present invention relates to a method for improving the nutrient uptake of a ruminant or ruminant-like animal comprising inoculating the animal with an immunogenic preparation effective in inhibiting the activity in the gastrointestinal tract of at least one species or strain of Archae. Preferably, the immunogenic preparation comprises an antigen component prepared from methanogenic bacteria.
Description
Method for Improving Utilisation of Nutrients by Ruminant or Ruminant-Like Animals FTFT.TD OF THE INVENTION
THIS INVENTION relates to methods for improving the utilisation of nutrients by ruminant and ruminant-like animals.
DISCUSSION OF THE PRIOR ART
The rumen or forestomach is an organ found in the digestive tract of certain herbivorous mammals prior to the gastric stomach and within which the digestion and formentation of plant material occurs through the activity of special microbial populations. A group of which some are found amongst such microbial populations are the Archae.
The Archae are a diverse group with regard to their physiology and include the methanogens, halophiles and thermacidophiles. They are related through ribosomal RNA
sequences that differ drastically from those of other procaryotes. Because of the anaerobic conditions in the rumen only those Archae that are obligate anaerobes are found therein. One member of the Archae found in the rumen, the methanogens are able to produce methane in symbiosis with other rumen microorganisms. For example, H2 and C02 are common end products and these may subsequently be used by the methanogens in the production of methane.
It has been recognised for some time, see e.g. Baxter, K.L.
(1962) 'The Energy Metabolism of ruminants' (Hutchinson, London), that rumen methanogenesis can result in a loss of energy available to an animal, a loss equivalent to possibly 10$ of an animal's gross energy intake. To reduce this loss several procedures for the inhibition of methanogenesis in the rumen have been investigated over the years. These procedures include the use of feed additives such as long-chain unsaturated fatty acids and the administration of ionophoric drugs to inhibit the production of Hz, Varying degrees of inhibition of methanogenesis have been demonstrated although in no case has it been clearly established that the overall result was economically favorable. Furthermore, irrespective of any technical inadequacies, the concept of constant administration of feed additives and drugs to meat-producing animals is increasingly regarded with disfavour by meat consumers.
BRIEF DESCRIPTION OF THE INVENTION
In accordance with a first aspect of the present invention there is provided a method for improving the nutrient uptake of a ruminant or fuminant-like animal comprising inoculating the animal with an immunogenic preparation effective in inhibiting the activity in the gastro-intestinal tract of at least one species or strain of Archae.
The ruminant animal may be a bovine, ovine, caprine, or cervine, while the ruminant-like animal may be a macropod, llama or alpaca.
The immunogenic preparation may comprise a whole cell preparation or a whole cell extract of one or more purified species or strains of Archae. Additionally, or alternatively, the preparation may comprise an immunogenic fraction (s), polypeptide (s) /peptide (s) polysaccharide (s) or lipolysaccharide (s) derived from one or more species or strains of Archae. Preferably the immunogenic preparation is effective in inhibiting the activity in the gastrointestinal tract of many species and strains of Archae, including the methanogenic bacteria.
THIS INVENTION relates to methods for improving the utilisation of nutrients by ruminant and ruminant-like animals.
DISCUSSION OF THE PRIOR ART
The rumen or forestomach is an organ found in the digestive tract of certain herbivorous mammals prior to the gastric stomach and within which the digestion and formentation of plant material occurs through the activity of special microbial populations. A group of which some are found amongst such microbial populations are the Archae.
The Archae are a diverse group with regard to their physiology and include the methanogens, halophiles and thermacidophiles. They are related through ribosomal RNA
sequences that differ drastically from those of other procaryotes. Because of the anaerobic conditions in the rumen only those Archae that are obligate anaerobes are found therein. One member of the Archae found in the rumen, the methanogens are able to produce methane in symbiosis with other rumen microorganisms. For example, H2 and C02 are common end products and these may subsequently be used by the methanogens in the production of methane.
It has been recognised for some time, see e.g. Baxter, K.L.
(1962) 'The Energy Metabolism of ruminants' (Hutchinson, London), that rumen methanogenesis can result in a loss of energy available to an animal, a loss equivalent to possibly 10$ of an animal's gross energy intake. To reduce this loss several procedures for the inhibition of methanogenesis in the rumen have been investigated over the years. These procedures include the use of feed additives such as long-chain unsaturated fatty acids and the administration of ionophoric drugs to inhibit the production of Hz, Varying degrees of inhibition of methanogenesis have been demonstrated although in no case has it been clearly established that the overall result was economically favorable. Furthermore, irrespective of any technical inadequacies, the concept of constant administration of feed additives and drugs to meat-producing animals is increasingly regarded with disfavour by meat consumers.
BRIEF DESCRIPTION OF THE INVENTION
In accordance with a first aspect of the present invention there is provided a method for improving the nutrient uptake of a ruminant or fuminant-like animal comprising inoculating the animal with an immunogenic preparation effective in inhibiting the activity in the gastro-intestinal tract of at least one species or strain of Archae.
The ruminant animal may be a bovine, ovine, caprine, or cervine, while the ruminant-like animal may be a macropod, llama or alpaca.
The immunogenic preparation may comprise a whole cell preparation or a whole cell extract of one or more purified species or strains of Archae. Additionally, or alternatively, the preparation may comprise an immunogenic fraction (s), polypeptide (s) /peptide (s) polysaccharide (s) or lipolysaccharide (s) derived from one or more species or strains of Archae. Preferably the immunogenic preparation is effective in inhibiting the activity in the gastrointestinal tract of many species and strains of Archae, including the methanogenic bacteria.
Preferably, the immunogenic preparation contains a whole cell preparation or whole cell extract of at least one species of methanogenic Archae.
Preferably, at least one methanogenic Archae is a member of a genus selected from the group consisting of Methanobacter, Methanobrevibacter and Methanosarcina.
Preferably, the methanogenic Archae is Methanobacter formicium.
Preferably, the whole cell preparation or whole cell extract is from a killed cell preparation.
Preferably, the killed cell preparation comprises a formaldehyde killed cell preparation.
The immunogenic preparation may further comprise one or more adjuvants of the type commonly used in the art, and may be provided as a veterinary preparation comprising an antigen component derived from one or more species or strains of Archae.
The immunogenic preparation or vaccine may be administered to the ruminant or ruminant-like animal via the intraperitoneal, intravenous or sub-cutaneous routes.
With regard to the method of the invention, the anaerobic fermentation of carbohydrates in the rumen results in the formation of volatile fatty acids, carbon dioxide and methane.
In this respect, it has been demonstrated that there is an inverse relationship between the production of methane and the production of propionate by rumen microorganisms. Indeed, wool production responds more to the supply of protein (which is spared by propionate) to the abomasums than to the supply of energy and, accordingly, the present invention may be expected to enhance wool growth in sheep.
Additionally, through a reduction in methane production the present invention may also provide benefits due to the belief that methane is a significant contributor to the "greenhouse"
effect.
In another aspect, the present inventions provides a method of inhibiting methane production in a ruminant or ruminant-like animal selected from the group consisting of macropod. Llama and alpaca comprising administering to the animal an immunogenic preparation containing a whole cell preparation or whole cell extract of at least one species of methanogenic Archae.
In another aspect, the present invention provides a method for improving the nutrient uptake of a ruminant or ruminant-like animal comprising inoculating the animal with an immunogenic preparation containing a whole cell preparation or whole cell extract of at least one species of methanogenic Archae.
In particular, the method of the present invention makes it possible for animals to have an improved productivity and nutrient uptake. In this respect, and as will be illustrated by the example below, animals to which the method is applied show improved characteristics, such as improved wool length growth (for sheep)and show an increased voluntary intake of dry matter and digestible dry matter, thereby increasing nutrient uptake.
These aspects of improved productivity and nutrient uptake are improvements over the normal characteristics of those same animals when not subjected to the method of the invention.
Preferably, at least one methanogenic Archae is a member of a genus selected from the group consisting of Methanobacter, Methanobrevibacter and Methanosarcina.
Preferably, the methanogenic Archae is Methanobacter formicium.
Preferably, the whole cell preparation or whole cell extract is from a killed cell preparation.
Preferably, the killed cell preparation comprises a formaldehyde killed cell preparation.
The immunogenic preparation may further comprise one or more adjuvants of the type commonly used in the art, and may be provided as a veterinary preparation comprising an antigen component derived from one or more species or strains of Archae.
The immunogenic preparation or vaccine may be administered to the ruminant or ruminant-like animal via the intraperitoneal, intravenous or sub-cutaneous routes.
With regard to the method of the invention, the anaerobic fermentation of carbohydrates in the rumen results in the formation of volatile fatty acids, carbon dioxide and methane.
In this respect, it has been demonstrated that there is an inverse relationship between the production of methane and the production of propionate by rumen microorganisms. Indeed, wool production responds more to the supply of protein (which is spared by propionate) to the abomasums than to the supply of energy and, accordingly, the present invention may be expected to enhance wool growth in sheep.
Additionally, through a reduction in methane production the present invention may also provide benefits due to the belief that methane is a significant contributor to the "greenhouse"
effect.
In another aspect, the present inventions provides a method of inhibiting methane production in a ruminant or ruminant-like animal selected from the group consisting of macropod. Llama and alpaca comprising administering to the animal an immunogenic preparation containing a whole cell preparation or whole cell extract of at least one species of methanogenic Archae.
In another aspect, the present invention provides a method for improving the nutrient uptake of a ruminant or ruminant-like animal comprising inoculating the animal with an immunogenic preparation containing a whole cell preparation or whole cell extract of at least one species of methanogenic Archae.
In particular, the method of the present invention makes it possible for animals to have an improved productivity and nutrient uptake. In this respect, and as will be illustrated by the example below, animals to which the method is applied show improved characteristics, such as improved wool length growth (for sheep)and show an increased voluntary intake of dry matter and digestible dry matter, thereby increasing nutrient uptake.
These aspects of improved productivity and nutrient uptake are improvements over the normal characteristics of those same animals when not subjected to the method of the invention.
21559 oa However, and with this in mind, because the Archae, including the methanogens, are naturally present in the rumen or forestomach of the vast majority of ruminant and ruminant-like animals it will be appreciated that animals exposed to the present invention are thus considered healthy animals and actually signify the normal population of any given animal species.
DESCRIPTION
The present invention will now be described, by way of example only, in relation to two examples. The examples illustrate various aspects of the present invention, but are in no way intended to limit the scope thereof.
EXAMPLE 1 Response of Sheep to the Intraperitoneal Administration of Antigen and Adjuvant An experiment was conducted using three test groups of ten (10) weaner wethers. The test groups are defined as follows:
Group I vaccine (antigen and adjuvant) administered intraperitoneally Group II adjuvant plus phosphate-buffered saline (PBS) administered intraperitoneally (control) Group III phosphate buffered saline (PBS) administered intraperitoneally (control) The vaccine was prepared from formaldehyde-killed cells of the following Archae:
DESCRIPTION
The present invention will now be described, by way of example only, in relation to two examples. The examples illustrate various aspects of the present invention, but are in no way intended to limit the scope thereof.
EXAMPLE 1 Response of Sheep to the Intraperitoneal Administration of Antigen and Adjuvant An experiment was conducted using three test groups of ten (10) weaner wethers. The test groups are defined as follows:
Group I vaccine (antigen and adjuvant) administered intraperitoneally Group II adjuvant plus phosphate-buffered saline (PBS) administered intraperitoneally (control) Group III phosphate buffered saline (PBS) administered intraperitoneally (control) The vaccine was prepared from formaldehyde-killed cells of the following Archae:
Methanobacter formicium (strain MF);
Methanobrevibacter arbor~hilus (strains AZ, DH1 and DC);
Methanobrevibacter ruminantium (strains M1 and Z6);
Methanobrevibacter smithii (strains AL1-A, B181 and PS); and Methanosarcina barberi (strain MS).
Approximately 2x109 cells of each strain were combined, washed and resuspended in 20m1 sterile phosphate-buffered saline (PBS) (pH7.2). Finally, an emulsion of lOml of this preparation (the antigen) and lOml Freund's complete adjuvant (FCA) was made, forming the vaccine used. The vaccine was administered (l.5ml per animal) as a primary inoculation intraperitoneally at the sub-lumbar fossa on the right mid-flank. The booster inoculation (1.5m1 per animal) was given subcutaneously twenty eight (28) days later, using the antigen preparation described above without added adjuvant (FCA).
The two control test groups were administered:
i) Freund's's complete adjuvant (FCA) emulsified in an equal volume of PBS (1.5m1 per animal) - Group II;
and ii) sterile PBS alone (l.5ml per animal) - Group III.
Each of i) and ii) were administered by the same route as the vaccine of Group I. The booster inoculation in each of test Groups II and III was sterile PBS alone (1.5m1 per animal) administered subcutaneously twenty eight (28) days after the primary inoculation.
Methanobrevibacter arbor~hilus (strains AZ, DH1 and DC);
Methanobrevibacter ruminantium (strains M1 and Z6);
Methanobrevibacter smithii (strains AL1-A, B181 and PS); and Methanosarcina barberi (strain MS).
Approximately 2x109 cells of each strain were combined, washed and resuspended in 20m1 sterile phosphate-buffered saline (PBS) (pH7.2). Finally, an emulsion of lOml of this preparation (the antigen) and lOml Freund's complete adjuvant (FCA) was made, forming the vaccine used. The vaccine was administered (l.5ml per animal) as a primary inoculation intraperitoneally at the sub-lumbar fossa on the right mid-flank. The booster inoculation (1.5m1 per animal) was given subcutaneously twenty eight (28) days later, using the antigen preparation described above without added adjuvant (FCA).
The two control test groups were administered:
i) Freund's's complete adjuvant (FCA) emulsified in an equal volume of PBS (1.5m1 per animal) - Group II;
and ii) sterile PBS alone (l.5ml per animal) - Group III.
Each of i) and ii) were administered by the same route as the vaccine of Group I. The booster inoculation in each of test Groups II and III was sterile PBS alone (1.5m1 per animal) administered subcutaneously twenty eight (28) days after the primary inoculation.
2t55908 The sheep were offered an ad lib:t~ ium diet comprised of 80~
oaten hay, 10~ oat grain, 7$ lupin grain, 1~ urea and 2$
mineral mix (sold under the brand name "Siromin").
Voluntary intake of dry matter was measured daily.
Live weight was measured every two weeks and the length growth rate of the wool was determined using dyebands applied at approximately four-week intervals. As noted above, the primary inoculation was followed by the second or booster inoculation 28 days later. The digestibility of the diet was measured before and after the treatments were administered (ie. before and after the sheep were inoculated).
The methane production of the microbial population from the rumen of each sheep was determined in vitro before and after each sheep was inoculated. An aliquot of rumen contents, obtained using a stomach tube, was incubated for 24 hours at 37°C in a culture broth of sterilised rumen liquor under an atmosphere of nitrogen and carbon dioxide. The methane content of the gases evolved during incubation was determined by gas chromatography.
The measurements made during the period prior to inoculation of the sheep were used as covariates in the statistical analysis of the measurements made after the sheep were inoculated.
Figure la illustrates the differences in the wool length growth rate (mm/day), between sheep in the above groups before and after inoculation. The wool length growth rate was greater in sheep given the vaccine (Group I) than in the control groups, for example the sheep given PBS alone (p<0.02).
Figure lb illustrates the differences in the dry matter intake (g/day), between sheep in the above groups before and after inoculation. Sheep inoculated with the vaccine (Group I) showed a significant increase in dry matter intake compared to those administered the adjuvant and PBS (Group II) (p<0.009).
Figure lc illustrates the differences in the digestible dry matter intake (g/day), between sheep in the above groups before and after inoculation. Inoculated sheep (Group 1) showed a significant increase in digestible dry matter intake (p<0.002) compared to those administered the adjuvant and PBS (Group II).
Figure ld illustrates the differences in methane production in vitro (ug/ml inoculum of rumen contents), between sheep in the above groups before and after inoculation. A
significant (p<0.018) reduction in methane production is evident in samples taken from sheep given the vaccine (Group I) compared with those samples taken prior to inoculation with the vaccine. In addition, there is a significantly lower production of methane in samples taken from sheep administered vaccine (Group I) compared with those samples taken from sheep in Group II or in Group III (P<0.018).
Figure le illustrates the differences in efficiency of wool production, expressed as length growth rate of wool (mm/d) relative to methane produced in vitro (ug/ml of rumen contents) in the above groups before and after inoculation.
A significant (p - 0.09) increase in efficiency of wool production is evident in sheep given the vaccine (Group 1) compared with those administered PBS alone (Group III).
Figure if illustratesthe differences methane production in in vitro(ug/ml) rumen contents) relative to digestible dry matter intake in the above groups before and after inoculation. A significant reduction (p < 0.06) in methane produced per 100g digestible dry matter intake (DDMI) is evident in sheep given the vaccines (Group I) compared with those administered the adjuvant and PBS (Group II).
EXAMPLE 2 Immunoglobulin Levels in Sheep after the Intraperitoneal Administration of an Antigen/
Adjuvant Vaccine.
An experiment was conducted using three test groups of eight (8) weaner wethers. Three vaccines were prepared, each using an antigen prepared from a washed preparation of mixed rumen protozoa and administered to a separate test group.
Frozen whole cells were thawed and 20m1 fractions (106 cells/ml) were incorporated into three vaccines as follows:
i) Freund's vaccine comprising 20m1 of the antigen preparation emulsified with 20m1 of Freund's Complete Adjuvant (FCA). The dose administered was 3ml per animal;
ii) Alum vaccine comprising 20m1 of the antigen preparation incorporated with "Alhydrogel" (Superfos, Denmark) according to the manufacturer's directions.
The dose administered was 2.5m1 per animal; and iii) DEAF-dextran sulphate vaccine comprising 20m1 of the antigen preparation, 20m1 of 20~ (w/v) diethlaminoethyl (DEAE)-dextran sulphate, lml of 30~
(v/v) glycerol, lml of 1~ (v/v) Tween 80 and 35m1 squalene mixed together. The mix formed an emulsion containing oil droplets no larger than 40um in diameter. The dose administered was 5ml per animal.
Each vaccine i), ii) and iii) was administered intraperitoneally at the sub-lumbar fossa on the right mid-flank of the animal for the primary inoculation. The booster inoculation in each case comprised a volume of the *Trade-mark _ g _ antigen preparation plus an equal volume of sterile PBS, being 1.5m1 administered subcutaneously 80 days after the primary inoculation.
A titre of antibody (IgG) was measured in both serum and saliva obtained from each animal on days 10, 21 and 113 after primary inoculation.
Figure 2a illustrates the significantly potentiated and sustained IgG levels (titre) in saliva obtained from animals administered the DEAE-dextran sulphate vaccine at each of the 10, 21 and 113 day samples compared with those of animals administered either vaccines i) or ii).
Figure 2b illustrates the significantly potentiated IgG
levels (titre) in serum obtained from animals administered with the DEAE-dextran sulphate vaccine at each of the 10, 21 and 113 day samples compared with those of animals administered either vaccines i) or ii).
It is evident from the above example that a DEAE-dextran and mineral oil, squalene or squalane emulsion when used as an immunoadjuvant together with an antigen prepared from washed rumen protozoa is capable of eliciting antibody titres approximately three (3) times greater than those obtained when FCA is used as the adjuvant in a vaccine administered to sheep (this represents a special or particular variation of the disclosure of International Application No. PCT/AU87/
00250 (WO 88/01177)). It is envisaged that similar results will be obtained if Archae, for example methanogens, are used as the antigen, or are used to produce the antigen.
Such results are considered significant because whilst FCA
is considered one of the most powerful immunoadjuvants presently available, it has not found wide application outside the laboratory because of the adverse tissue reaction it provokes in recipient animals, including the production of granulomateous lesions at the site of inoculation.
Accordingly, it is expected that the immunogenic activity of a vaccine produced from Archae and a combination of DEAE-dextran and an immunoadjuvant oil such as a mineral oil, squalane or squalene will be similarly effective in generating antibody titres that compare with those elicited when Archae and FCA are used as the antigen/adjuvant combination in the preparation of a vaccine. As such, it is envisaged that given such a immunogenic activity in such a vaccine there will be a significant inhibition of activity in the gastro-intestinal tract of one or more species or strains of Archae, dependent upon which species or strains were used to prepare the antigen employed in the vaccine, and which does not have the adverse reactions seen when FCA
is used.
It is further envisaged that similarly significant levels of the growth rate of wool, dry matter intake and digestible dry matter intake may be achieved when compared with the levels demonstrated using a FCA vaccine in Example 1.
It is still further envisaged that the present invention, through the inhibition of methane production, provides the opportunity to (i) manipulate body composition of ruminants by improving the balance of protein and energy supplied to an animal; and (ii) provide leaner carcasses in lot-fed animals.
Modifications and variations such as would be apparent to the skilled addressee upon a reading of this specification are considered to fall within the scope of the present invention.
oaten hay, 10~ oat grain, 7$ lupin grain, 1~ urea and 2$
mineral mix (sold under the brand name "Siromin").
Voluntary intake of dry matter was measured daily.
Live weight was measured every two weeks and the length growth rate of the wool was determined using dyebands applied at approximately four-week intervals. As noted above, the primary inoculation was followed by the second or booster inoculation 28 days later. The digestibility of the diet was measured before and after the treatments were administered (ie. before and after the sheep were inoculated).
The methane production of the microbial population from the rumen of each sheep was determined in vitro before and after each sheep was inoculated. An aliquot of rumen contents, obtained using a stomach tube, was incubated for 24 hours at 37°C in a culture broth of sterilised rumen liquor under an atmosphere of nitrogen and carbon dioxide. The methane content of the gases evolved during incubation was determined by gas chromatography.
The measurements made during the period prior to inoculation of the sheep were used as covariates in the statistical analysis of the measurements made after the sheep were inoculated.
Figure la illustrates the differences in the wool length growth rate (mm/day), between sheep in the above groups before and after inoculation. The wool length growth rate was greater in sheep given the vaccine (Group I) than in the control groups, for example the sheep given PBS alone (p<0.02).
Figure lb illustrates the differences in the dry matter intake (g/day), between sheep in the above groups before and after inoculation. Sheep inoculated with the vaccine (Group I) showed a significant increase in dry matter intake compared to those administered the adjuvant and PBS (Group II) (p<0.009).
Figure lc illustrates the differences in the digestible dry matter intake (g/day), between sheep in the above groups before and after inoculation. Inoculated sheep (Group 1) showed a significant increase in digestible dry matter intake (p<0.002) compared to those administered the adjuvant and PBS (Group II).
Figure ld illustrates the differences in methane production in vitro (ug/ml inoculum of rumen contents), between sheep in the above groups before and after inoculation. A
significant (p<0.018) reduction in methane production is evident in samples taken from sheep given the vaccine (Group I) compared with those samples taken prior to inoculation with the vaccine. In addition, there is a significantly lower production of methane in samples taken from sheep administered vaccine (Group I) compared with those samples taken from sheep in Group II or in Group III (P<0.018).
Figure le illustrates the differences in efficiency of wool production, expressed as length growth rate of wool (mm/d) relative to methane produced in vitro (ug/ml of rumen contents) in the above groups before and after inoculation.
A significant (p - 0.09) increase in efficiency of wool production is evident in sheep given the vaccine (Group 1) compared with those administered PBS alone (Group III).
Figure if illustratesthe differences methane production in in vitro(ug/ml) rumen contents) relative to digestible dry matter intake in the above groups before and after inoculation. A significant reduction (p < 0.06) in methane produced per 100g digestible dry matter intake (DDMI) is evident in sheep given the vaccines (Group I) compared with those administered the adjuvant and PBS (Group II).
EXAMPLE 2 Immunoglobulin Levels in Sheep after the Intraperitoneal Administration of an Antigen/
Adjuvant Vaccine.
An experiment was conducted using three test groups of eight (8) weaner wethers. Three vaccines were prepared, each using an antigen prepared from a washed preparation of mixed rumen protozoa and administered to a separate test group.
Frozen whole cells were thawed and 20m1 fractions (106 cells/ml) were incorporated into three vaccines as follows:
i) Freund's vaccine comprising 20m1 of the antigen preparation emulsified with 20m1 of Freund's Complete Adjuvant (FCA). The dose administered was 3ml per animal;
ii) Alum vaccine comprising 20m1 of the antigen preparation incorporated with "Alhydrogel" (Superfos, Denmark) according to the manufacturer's directions.
The dose administered was 2.5m1 per animal; and iii) DEAF-dextran sulphate vaccine comprising 20m1 of the antigen preparation, 20m1 of 20~ (w/v) diethlaminoethyl (DEAE)-dextran sulphate, lml of 30~
(v/v) glycerol, lml of 1~ (v/v) Tween 80 and 35m1 squalene mixed together. The mix formed an emulsion containing oil droplets no larger than 40um in diameter. The dose administered was 5ml per animal.
Each vaccine i), ii) and iii) was administered intraperitoneally at the sub-lumbar fossa on the right mid-flank of the animal for the primary inoculation. The booster inoculation in each case comprised a volume of the *Trade-mark _ g _ antigen preparation plus an equal volume of sterile PBS, being 1.5m1 administered subcutaneously 80 days after the primary inoculation.
A titre of antibody (IgG) was measured in both serum and saliva obtained from each animal on days 10, 21 and 113 after primary inoculation.
Figure 2a illustrates the significantly potentiated and sustained IgG levels (titre) in saliva obtained from animals administered the DEAE-dextran sulphate vaccine at each of the 10, 21 and 113 day samples compared with those of animals administered either vaccines i) or ii).
Figure 2b illustrates the significantly potentiated IgG
levels (titre) in serum obtained from animals administered with the DEAE-dextran sulphate vaccine at each of the 10, 21 and 113 day samples compared with those of animals administered either vaccines i) or ii).
It is evident from the above example that a DEAE-dextran and mineral oil, squalene or squalane emulsion when used as an immunoadjuvant together with an antigen prepared from washed rumen protozoa is capable of eliciting antibody titres approximately three (3) times greater than those obtained when FCA is used as the adjuvant in a vaccine administered to sheep (this represents a special or particular variation of the disclosure of International Application No. PCT/AU87/
00250 (WO 88/01177)). It is envisaged that similar results will be obtained if Archae, for example methanogens, are used as the antigen, or are used to produce the antigen.
Such results are considered significant because whilst FCA
is considered one of the most powerful immunoadjuvants presently available, it has not found wide application outside the laboratory because of the adverse tissue reaction it provokes in recipient animals, including the production of granulomateous lesions at the site of inoculation.
Accordingly, it is expected that the immunogenic activity of a vaccine produced from Archae and a combination of DEAE-dextran and an immunoadjuvant oil such as a mineral oil, squalane or squalene will be similarly effective in generating antibody titres that compare with those elicited when Archae and FCA are used as the antigen/adjuvant combination in the preparation of a vaccine. As such, it is envisaged that given such a immunogenic activity in such a vaccine there will be a significant inhibition of activity in the gastro-intestinal tract of one or more species or strains of Archae, dependent upon which species or strains were used to prepare the antigen employed in the vaccine, and which does not have the adverse reactions seen when FCA
is used.
It is further envisaged that similarly significant levels of the growth rate of wool, dry matter intake and digestible dry matter intake may be achieved when compared with the levels demonstrated using a FCA vaccine in Example 1.
It is still further envisaged that the present invention, through the inhibition of methane production, provides the opportunity to (i) manipulate body composition of ruminants by improving the balance of protein and energy supplied to an animal; and (ii) provide leaner carcasses in lot-fed animals.
Modifications and variations such as would be apparent to the skilled addressee upon a reading of this specification are considered to fall within the scope of the present invention.
Claims (21)
1. A method for improving the nutrient uptake of a ruminant or ruminant-like animal comprising inoculating the animal with an immunogenic preparation containing a whole cell preparation or whole cell extract of at least one species of methanogenic Archae.
2. The method according to claim 1, wherein the at least one species of methanogenic Achae is a member of a genus selected from the group consisting of Methanobacter, Methanobrevibacter and Methanosarcina.
3. The method according to claim 1, wherein the at least one species of methanogenic Archae is selected from the group consisting of Methanobacter formicium, Methanobrevibacter arboriphilus, Methanobrevibacter ruminatium, Methanobrevibacter smithil and Methanosarcina barkeri.
4. The method according to claim 1, wherein the methanogenic Archae is selected from the group consisting of Methanobacter formicium, Methanobrevibacter ruminantium and Methanoscarcina bakeri.
5. The method according to claim 1, wherein the methanogenic Archae is Methanobacter formicium.
6. The method according to any one of claims 1 to 5, wherein the immunogenic preparation further comprises an adjuvant.
7. The method according to any one of claims 1 to 6, wherein the immunogenic preparation is administered via the intraperitoneal route.
8. The method according to any one of claims 1 to 6, wherein the inoculation of the animal comprises a primary inoculation with a immunogenic preparation administered via the intraperitoneal route and a secondary or booster inoculation with the same or different immunogenic preparation administered some time after.
9. The method according to claim 8, wherein the booster inoculation is with the immunogenic preparation containing a whole cell preparation or whole cell extract of at least one species of methanogenic Archae.
10. The method according to any one of claims 6 to 9, wherein the adjuvant contains DEAE-dextran.
11. The method according to any one of claims 6 to 9, wherein the adjuvant contains diethylaminoethyl (DEAE)-dextran and an immunoadjuvant oil.
12. The method according to claim 11, wherein the immunoadjuvant oil is selected from the group consisting of a mineral oil, squalene and squalane.
13. The method according to any one of claims 1 to 12, wherein the inoculation with the immunogenic preparation provides a significant increase in the efficiency of utilisation of nutrients upon an inhibition of methane production.
14. The method according to claim 13, wherein the increased efficiency includes a significant increase in the length growth rate of wool relative to methane production.
15. The method according to claim 13, wherein the increased efficiency includes a significant decrease in methane production relative to digestible dry matter intake.
16. The method according to any one of claims 1 to 15, wherein said ruminant animal is selected from the group consisting of bovine, ovine, caprine, and cervine.
17. The method according to any one of claims 1 to 15, wherein said ruminant-like animal is selected from the group consisting of macropod, llama and alpaca.
18. The method according to claim 1, wherein said immunogenic preparation contains a whole cell preparation or whole cell extract from a plurality of species of methanogenic Archae.
19. The method according to claim 1, wherein the whole cell preparation or whole cell extract is from a killed cell preparation.
20. The method according to claim 19, wherein the killed cell preparation comprises a formaldehyde killed cell preparation.
21. A method for inhibiting methane production in a ruminant or ruminant-like animal selected from the group consisting of macropod, llama and alpaca comprising administering to the animal an immunogenic preparation containing a whole cell preparation or whole cell extract of at least one species of methanogenic Archae.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2155908 CA2155908C (en) | 1995-08-11 | 1995-08-11 | Method for improving utilisation of nutrients by ruminant or ruminant-like animals |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2155908 CA2155908C (en) | 1995-08-11 | 1995-08-11 | Method for improving utilisation of nutrients by ruminant or ruminant-like animals |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2155908A1 CA2155908A1 (en) | 1997-02-12 |
| CA2155908C true CA2155908C (en) | 2005-08-09 |
Family
ID=4156409
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2155908 Expired - Fee Related CA2155908C (en) | 1995-08-11 | 1995-08-11 | Method for improving utilisation of nutrients by ruminant or ruminant-like animals |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2155908C (en) |
-
1995
- 1995-08-11 CA CA 2155908 patent/CA2155908C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CA2155908A1 (en) | 1997-02-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6036950A (en) | Method for improving utilisation of nutrients by ruminant or ruminant-like animals | |
| US5589168A (en) | Probiotic | |
| EP0831804A1 (en) | Methods for maintaining or elevating the cd-4 or cd-8 cell levels comprising the administration of conjugated linoleic acid; a new process for the manufacture of conjugated linoleic acid using lactobacillus sp | |
| CN101094685B (en) | The purposes of flagellin in the immunotherapy of Yersinia pestis | |
| Moreau et al. | Hydrolysis of urea in the gastrointestinal tract of" monoxenic" rats: effect of immunization with strains of ureolytic bacteria | |
| EP0063490B1 (en) | Lactation improvement method | |
| AU2003281529B2 (en) | Composition for promoting the proliferation of lactobacillus casei subsp. casei | |
| AU692481B2 (en) | Method for improving utilisation of nutrients by ruminant or ruminant-like animals | |
| CA2155908C (en) | Method for improving utilisation of nutrients by ruminant or ruminant-like animals | |
| US5176910A (en) | Treponema hyodysenteriae hemolysin and uses therefor | |
| Jensen et al. | The characteristics of some rumen lactobacilli | |
| CN1033008A (en) | anti-poultry E. coli septicemia vaccine | |
| US6689354B2 (en) | Immunogenic preparation and method for improving the productivity of ruminant animals | |
| CN108904791A (en) | A kind of clostridium perfringens alpha toxin recombinant subunit vaccine and its production method | |
| US6287555B1 (en) | Method and vaccine for prevention of over-production of acid in the rumen or gut of animals | |
| RU2291194C2 (en) | Consortium of bifidobacterium strains used in preparing liquid concentrate of bifidobacterium, fermented-milk, nonfermented, curative-prophylactic foodstuffs, biologically active supplements and bacterial preparations | |
| CA1276084C (en) | Composition and method for improving the efficiency of ruminant feed utilization | |
| CA3068257A1 (en) | Compositions and methods for reducing methane producing bacteria in animals | |
| KR100206454B1 (en) | A novel lactobacillus sp ds-12 and use as a probiotic for fish | |
| JPH06505226A (en) | Reduced protein subunit vaccine for swine dysentery | |
| RU2173159C2 (en) | Method of production of biologically active food addition | |
| WO2009051511A1 (en) | Method for producing a liquid or dried bifidobacteria concentrate or a starter | |
| RU2232589C2 (en) | Method for obtaining probiotic additive for fodder means | |
| JPH11507377A (en) | ACTINOMYCES PYOGENES and ACTINOMYCES PYOGENES / FUSOBACTERIUM NECROPHORUM Vaccines for Bovine and Sheep Immunization | |
| CN116649465A (en) | Feed additive produced by using bacillus subtilis and production method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |