CA1337971C

CA1337971C - Products and processes for increasing the growth rate and /or utilization of feeds or foods and/or resistance in animals and humans

Info

Publication number: CA1337971C
Application number: CA000579451A
Authority: CA
Inventors: Reinhard Teichmann; Hans-Georg Liebich
Original assignee: Effem GmbH
Current assignee: Mars GmbH
Priority date: 1987-10-07
Filing date: 1988-10-06
Publication date: 1996-01-23
Anticipated expiration: 2013-01-23
Also published as: DE3733899A1; DE3733899C2

Abstract

The present invention is a product and process for increasing the growth rate and/or, feed or food utilization and/or resistance in animals and humans characterized by at least one antigen for the immuniza-tion of animals and humans and prior and/or simultaneous and/or subsequent gastrointestinal administration, and at least one immunostimulating or immunodepressing agent.

Description

30,963 1~37371 PRODUCTS AND PROCESSES FOR INCREASING THE GROWTH
RATE AND/OR UTILIZATION OF FEEDS OR FOODS AND/OR
RESISTANCE IN ANIMALS AND HUMANS
Feeds and feed additives have been used for many decades, and other processes for promoting animal growth and for improving nutrient utilization have had limited success.
Antibiotics in feeds may not only lead to irrepar-able damage to animal health through the increasing formation of resistant pathogen strains with the danger of secondary diseases, but also may be dangerous to the health of humans, as the final consumers of the foods of animal origin.
Hormones may cause even greater damage to animal and human health, among other reasons, because of their anabolic promotion of the growth rate. Furthermore, the above-mentioned active substances may not be ex-creted from the animal body to the desired extent, resulting in certain residue problems.
Secretion and thus digestion in the stomach are customarily divided into the cephalic, gastric, and intestinal phases. The secretion of acid and pepsin is stimulated to the greatest extent in the gastric phase.
During this phase, the antral hormone gastrin is re-leased as the most important stimulus for acid secre-tion. Acid secretion is necessary for denaturation of the proteins and their degradation, and is also the ~' first infection barrier. The acid, which passes through the pylorus into the duodenal bulb, leads to the release of secretin and also, to a small extent, of cholecystokinin. Secretin strongly stimulates the aqueous components of the pancreatic secretion and thus flushes out the pancreatic enzymes. Through these mechanisms, the whole cascade of digestion is initiated.
A key hormone in the gastric phase of digestion and, consequently, for induction of the digestive processes in the gastrointestinal tract is gastrin. Although this is released through neurogenic, mechanical, and hormonal stimuli, this release is, however, decisively dependent on the individual food components passing into the stomach. Proteins and peptides are the important substances in the food that release gastrin.
The present invention starts from the finding that, with each meal, the mucosa of the stomach comes into contact with substances that could show immunobiological activity as antigens in the gastrointestinal tract.
The advantage of the present invention is therefore to indicate products and processes for increasing the growth rate and/or the utilization of feeds and foods and/or the resistance in animals and humans, which are not only biologically effective but also free of any residue problems.
This invention relates to a gastrointestinal use of a pharmaceutically effective amount of an antigen in an animal immunized against said antigen for increasing the growth rate or food utilization or for increasing resistance in said animal.
The invention further relates to a commercial package comprising an immunologically effective amount of an antigen ~B~

2a 69130-11 together with instructions for use thereof (a) to immunize an animal and (b) to apply the antigen gastrointestinally to increase the growth rate of said animal or to increase immunological resistance of said animal.
The invention also relates to an agricultural package comprising an immunologically effective amount of an antigen in forms suitable (a) to immunize said animal by injection and ~b) apply to said animal gastrointestinally.
Particular preferred embodiments of the invention are those:
(a) wherein the antigen is selected from the group consisting of amino acids, peptides, proteins, scleroproteins, globular proteins, histones, albumins, globulins, proteids, nucleic acids, nucleosides, nucleotides, porphyrins, cytochromes, lipids, phospholipids, glycolipids, lipoproteins, fatty acids, cerebrosides, gangliosides, isoprenoid lipids, steroids, carotinoids, polyprenols, polyprenylquinones, monosaccharides, glycosides, oligosaccharides, glycogen, homoglycanes, heteroglycanes, glycoproteins, microorganisms and protozoa;
(b) wherein the antigen is a catabolic or metabolic product;
(c) wherein the antigen is a synthetic peptide, selected from the group consisting of homopolymers, statistical or alternating copolymers, block polymers and graft copolymers, with antigenic determinants for stimulating an immune response;
(d) wherein the antigen is hapten, selected from the group consisting of nitrophenylacetic acid and 2,4-dinitrofluorobenzene;
(e) wherein the antigen is a hapten coupled to a carrier protein;
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2b 69130-11 (f) wherein the antigen is in unit dose form suitable for application in an enteral manner in a quantity of 1 to 500 mg/kg of body weight;
(g) wherein the antigen is in unit dose form suitable for immunization in a quantity of 1 to 500 mg/kg of body weight;
(h) wherein the antigen is in unit dose form suitable for parenteral administration in a quantity of 1 to 100 mg/kg of body weight.
The antigen may be capable of being applied before, during or after immunization.
The present invention is surprisingly effective given increase of the growth rate and/or utilization of feeds and foods in amounts of at least several tens of percent, and is achieved, as has been demonstrated by the inventors particularly in pig fattening (compare Experiments 1 and 2 below) by means of an increase of 30.4% in feed utilization in that case.

Investigations on Doqs In non-immunized dogs (20 mongrel dogs), the administration into the stomach of an antigen, namely, human gamma-globulin (HGG), as a protein foreign to the dog, did not produce any hormonal stimulation.
In immunized dogs, on the other hand, there was a highly significant and large release of serum gastrin, simultaneously with a significant increase in the blood flow in the mucosa of the antrum, the duodenal bulb, and the duodenal mucosa after ten minutes.
After the intragastric administration of human albumin as a non-antigenic protein, no release of gastrin was found, which proves the specificity of the immunological reaction.
When the degree of immunization decreased, approximately eight weeks after the last booster dose, the administration of HGG to the canine stomach produced only a slight release of gastrin. Reimmuniza-tion again led to a significant gastrin increase.
It was shown, additionally, that the feeding of commercial dog food before and after the immunization with HGG did not produce any changes in the gastrin release.
The gastrin release was also compared with that after a known protein stimulation, such as commercial formula diet for enteral human feeding (Biosorbin~) with a 5-gram protein component, dissolved in an equal volume. This produced practically no hormone release, although the protein and peptide content, calculated on a molar basis, is higher in Biosorbin~ than in the HGG
solution.
On the other hand, the administration of 5 grams of bovine serum albumin (BSA), a protein known to the dog, led to a significant stimulation corresponding to that after dog food.
The secretin release was also compared.
A strong stimulation of secretin occurred after the administration of one gram of HGG to immunized animals, just as strong as after a food stimulation.
However, no secretin release took place after the administration of BSA, probably as a result of the high buffer capacity of this substance.
Cardiovascular and respiratory parameters:
There were no significant changes in the cardiac output, blood pressure, or blood gases in both groups of animals during the study, i.e., neither in immunized nor non-immunized animals with and without antigen administration.
No effects on the respiratory functions were observed.

Investiqations on Rats The phenomenon of immunological stimulation of digestion discovered according to the invention was also demonstrated in rats.

Application of the Investigations The improved digestion and metabolism of the protein used as an antigen ~and of all proteins in the food~ depends on the pancreatic enzymes.
This phenomenon can be explained by the fact that the whole gastroenteropancreatic cascade can be immunologically stimulated:
- it starts with the gastrin release, 13~7971 - followed by the production of acid and mucus, - which finally leads to a secretin release with all of its effects.
The discovery of the immunological mechanism for stimulation of the digestive functions represents a new and important contribution for effective control of the digestion.
This mechanism uses physiological processes; it is "more biological", its effectiveness has been demon-strated in field tests under economically relevant conditions.
An improved feed or food utilization is of maximum importance for animals and humans with a similarly functioning gastrointestinal system, which reacts similarly to this immunological mechanism discovered according to the invention. The identical reaction in the dog, rat, and pig proves this.
The outstanding characteristic of the mechanism discovered, on which the invention is based, is the greatly increased digestive capacity through the induction of, primarily, gastric and, in particular, pancreatic secretion.
If small quantities of the antigen against which the animal had been immunized pass into the stomach, these represent a very strong stimulus of gastroin-testinal functions. Gastrin and secretin are then released, and the gastric and pancreatic secretion increases. These processes show that small quantities of, for example, a protein in animals immunized against this induce a large increase in digestive capacity.
What is decisive for the stimulation of the digestion through these immunological mechanisms is not the quantity of food ingested, but the composition of the food. The food must, for example, contain the protein that the stomach can recognize immunologically.
This immunological stimulus is even greater than that triggered by the vagus nerve, hormones, or dilation of the stomach. These discoveries by the inventors about the immunological mechanism are the scientific basis of the growth promotion and the improvement of the feed and food utilization.
The treatment consequently includes the use of a naturally occurring or synthetic substance, which is either already present in the feed or in human food or can be added, provided that the animal or human is sensitized against this special substance. An oral sensitization takes place predominantly only in a newborn or premature offspring. The treatment consists of administering a biological (i.e., naturally occurr-ing) or synthetic substance (or hapten), against which the organism is immunized, in the feed or food, in the drinking water or other fluids, in solid or liquid form as a tablet, bolus as granules, or in juices, syrup, or molasses or sauces. In addition, the material can be added alone or in combination with other already tested or generally recognized as safe substances or foods that are used routinely for similar or other health purposes or the like.
The quantity of biologically active additive, i.e., for example, protein or hapten, as antigen that is required depends on the:
- composition of the feed or food, - species, - age, - sex, and - breeding of treated individuals or populations, - dose and duration of the immunization.
The following empirical dose is given as a guideline:

1 gram of protein in 100 grams of liquid feed for intragastric stimulation.
The presence of antibodies from immunizations is a requirement for the effectiveness of the intragastric antigen administration.

Aqents with Immunostimulatinq or Immunosuppressive Action There are groups of biological and synthetic substances, and also products of activated cells, that have a stimulating or suppressive effect on the immune system.
These agents act via T-lymphocytes, B-lymphocytes, macrophages, or natural killer cells.
The agents can be given alone or in combination with agents to promote growth, feed utilization and resistance.
Some agents of this type have already received successful clinical use, particularly the immunostimulating agents - BCG in mastocarcinomas, melanomas, lymphosarcomas, and bronchial carcinomas;
- Bestatin in gastric carcinomas and mastocarcinomas, - Interferons in hepatitis-B, herpes zoster and cytomegalic disease, - Thymus hormones in immunodeficiencies; the immunosuppressive agent - Cyclosporin, frequently in organ transplants.
Other agents of this type have been tested at least in vitro.
Optimum results are achieved in the fattening of animals for meat production, namely a shortening of the fattening time, with smaller feed consumption to reach the final carcass weight.

In addition, this immunobiologically active, growth-promoting effect promotes the resistance mechanisms in the body, particularly in the case of stress, through specific and non-specifically active defense systems of peripheral lymphatic organs and circulating lymphocytes in the blood and lymph.

Experiment 1 (pilot experiment) In this preliminary experiment concluded under field conditions, piglets after weaning from the mother animals were immunized with whey protein in a pig fattening operation. After this, the animals were given whey, added to the drinking water.
Although all of the animals (n = 20) were suffer-ing from an infectious intestinal disease, growth of the animals was hindered by the infection only in the ten untreated control animals. After disappearance of the infection, an even higher growth rate started in the treated animals.
The daily weight gain up to the carcass weight was 675 grams in the treated group and 600 grams in the untreated group.
The feed utilization, i.e., the amount of feed per kilogram required to increase the body weight by one kilogram, was 1:2.45 in the treated group, as compared with 1:3.25 in the controls.
In this pilot study, the daily weight gain improv-ed by ~12.5% and the feed utilization by ~24%.
These experiences and observations formed the basis for a second series of experiments.

Experiment 2 (double blind experiment) Duration: 25 weeks Experimental Animals: 44 pigs:
improved German Landrace pig, 22 male and 22 female animals, divided into four groups of 11 animals each as piglets.
Pretreatment and Feeding: The piglets were weaned from the mother animals at the age of 28 days Up to week 10 the animals received commercial, stand-ardized piglet rearing feed.
After weaning, the animals were assigned to one of the following groups on a random basis:
Group 1: parenteral sensitization with whey and individual weighing.
Group 2: parenteral sensitization with whey and determination of the group weight at the beginning and end of the experiment.
Group 3: no immunization (saline injection) and individual weighing.
Group 4: No immunization (saline injection) and determina-tion of the group weight at the beginning and end of the experiment.

Sensitization:
The animals of Groups 1 and 2 were immunized three times, at the age of 4, 7 and 10 weeks, by subcutaneous injection of l.O mL of commercial, concentrated milk-protein solution (whey, protein content 10%).
From week 10 on, the animals received a commercial fattening feed of uniform composition and also were given whey in the drinking water. In this way, a continuous enterogastric stimulation between the orally ingested milk protein and the parenterally sensitized gastric mucosa was ensured.

Procedure:
Groups of 11 pigs were housed in a shed with a slatted floor without litter, with liquid feeding twice a day (whey and fattening feed) and free drinking (whey in the drinking water). The amount of feed consumed and amount of whey were determined and recorded electronically for the individual boxes.

Weiqhing of the Animals:
All animals were weighed as a group at the beginning of the experiment, after ten weeks, and at the end of the experiment after 15 weeks. In Groups 1 and 3, all animals were also weighed individually, at the end of weeks 3, 5, 7, and 11 of the fattening period. The average weight per animal and group was determined.

Slauqhtering and Determination of the Meat Ouality:
All animals were slaughtered after 15 weeks (at the age of approximately 25 weeks) and the meat was classified according to the EC guidelines.

1~37971 Results The daily weight gain, total body weight at the end of the experiment, and weights and feed utilization rates of all groups are shown in Table I and Figures 1 and 2.
The evaluation of the carcasses animals, inde-pendent of the treatment, produced the classification E-l (excellent) according to the EC guidelines.
The results of Experiment 2 show clearly that, after previous immunization and subsequent entero-gastric antigen administration, the feed utilization, and thus the daily weight gain, increased considerably.
The live and carcass weight of the immunized animals are therefore increased significantly after 15 weeks, by ~16% and ~18.5%, respectively. These results correspond to those of Experiment 1 and show clearly that the treatment, i.e., immunization and (subsequent) administration of the antigen with the feed, for animals under otherwise identical conditions, is not only extremely effective with an increase of 30.4% in feed utilization.
but is also extremely advantageous economically for the breeder, since the feed consumption for fattening decreases distinctly by 13.5%.
No side effects were found with this treatment.
On the contrary, the pigs in Groups 1 and 2 recovered from a viral infection, which caused diarrhea, more rapidly than the control animals; the daily weight gain, after the infection had been overcome, immediately started to increase again (Figure 2), but not in the control group.
These results and investigations, which were carried out under field conditions in both experiments, clearly proved that the invention - on the basis of -l2- 1337971 investigations on laboratory animals - can be used successfully in animal production.

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Claims

1. A gastrointestinal use of a growth rate, food utilization or resistance increasing effective amount of an antigen in an animal immunized against said antigen for in-creasing the growth rate or food utilization or for increasing resistance in said animal.

2. The use according to claim 1, wherein the antigen is selected from the group consisting of amino acids, peptides, proteins, scleroproteins, globular proteins, histones, albumins, globulins, proteids, nucleic acids, nucleosides, nucleotides, porphyrins, cytochromes, lipids, phospholipids, glycolipids, lipoproteins, fatty acids, cerebrosides, gangliosides, iso-prenoid lipids, steroids, carotinoids, polyprenols, polyprenyl-quinones, monosaccharides, glycosides, oligosaccharides, glycogen, homoglycanes, heteroglycanes, glycoproteins, micro-organisms and protozoa.

3. The use according to claim 2, wherein the antigen is a catabolic or metabolic product.

4. The use according to claim 1, wherein the antigen is a synthetic peptide, selected from the group consisting of homo-polymers, statistical or alternating copolymers, block polymers and graft copolymers, with antigenic determinants for stimulat-ing an immune response.

5. The use according to claim 4, wherein the antigen is hapten, selected from the group consisting of nitrophenylacetic acid and 2,4-dinitrofluorobenzene.

6. The use according to claim 1, wherein the antigen is a hapten coupled to a carrier protein.

7. The use of claim 1, wherein the antigen is capable of being applied gastrointestinally prior to immunizing the animal with the antigen.

8. The use of claim 1, further wherein the antigen is capable of being applied gastrointestinally simultaneously with immunizing the animal with the antigen.

9. Use of a growth rate, food utilization or resistance increasing effective amount of an antigen gastrointestinally after immunization by the antigen to increase the growth rate or food utilization or to increase resistance in an animal.

10. The use according to claim 9, wherein the antigen is selected from the group consisting of amino acids, peptides, proteins, scleroproteins, globular proteins, histones, albumins, globulins, proteids, nucleic acids, nucleosides, nucleotides, porphyrins, cytochromes, lipids, phospholipids, glycolipids, lipoproteins, fatty acids, cerebrosides, gangliosides, iso-prenoid lipids, steroids, carotinoids, polyprenols, polyprenyl-quinones, monosaccharides, glycosides, oligosaccharides, polysaccharides, glycogen, homoglycanes, heteroglycanes, glycoproteins, microorganisms and protozoa.

11. The use according to claim 10, wherein the antigen is a catabolic or metabolic product.

12. The use according to claim 9, wherein the antigen is a synthetic peptide, selected from the group consisting of homopolymers, statistical or alternating copolymers, block polymers and graft copolymers, with antigenic determinants for stimulating an immune response.

13. The use according to claim 12, wherein the antigen is a hapten, selected from the group consisting of nitrophenylacetic acid and 2,4-dinitrofluorobenzene.

14. The use according to claim 9, wherein the antigen is a hapten coupled to a carrier protein.

15. The use according to claim 9, wherein the immunization is achieved by a quantity of antigen of 1 to 500 mg/kg of body weight.

16. The use according to claim 15, wherein the immunization is achieved by administration of the antigen once or several times parenterally in a quantity of 1 to 100 mg/kg of body weight.

17 17. The use according to claim 9, wherein the antigen is in a form suitable for daily application and in enteral manner in a quantity of 2 to 500 mg/kg of body weight.

18. The use of claim 9 wherein the antigen is whey.

19. The use of claim 18 wherein the animal is a pig.

20. The use of claim 9 wherein the animal is a dog.

21. The use of claim 9 wherein the animal is a rat.

22. The use of claim 1 or 9 for simultaneously increasing the growth rate, food utilization and immunological resistance in an animal.

23. A commercial package comprising an immunologically effective amount of an antigen together with instructions for use thereof (a) to immunize an animal and (b) to apply the antigen gastrointestinally to increase the growth rate of said animal or to increase immunological resistance of said animal.

24. An agricultural package comprising an immunologically effective amount of an antigen in forms suitable (a) to immunize said animal by injection and (b) apply to said animal gastrointestinally.

25. A commercial package according to claim 23, wherein the antigen is selected from the group consisting of amino acids, peptides, proteins, scleroproteins, globular proteins, histones, albumins, globulins, proteids, nucleic acids, nucleosides, nucleotides, porphyrins, cytochromes, lipids, phospholipids, glycolipids, lipoproteins, fatty acids, cerebrosides, gangliosides, isoprenoid lipids, steroids, carotinoids, polyprenols, polyprenylquinones, monosaccharides, glycosides, oligosaccharides, glycogen, homoglycanes, heteroglycanes, glycoproteins, microorganisms and protozoa.

26. A commercial package according to claim 23, wherein the antigen is a synthetic peptide, selected from the group consisting of homopolymers, statistical or alternating copolymers, block polymers and graft copolymers, with antigenic determinants for stimulating an immune response.

27. A commercial package according to claim 26, wherein the antigen is hapten, selected from the group consisting of nitrophenylacetic acid and 2,4-dinitrofluorobenzene.

28. A commercial package according to claim 23, wherein the antigen is a hapten coupled to a carrier protein.

29. A commercial package according to claim 23, wherein the antigen is in a form capable of being applied gastro-intestinally prior to immunization of said animal with the antigen.

30. A commercial package according to claim 23! wherein the antigen is in a form capable of being applied gastro-intestinally simultaneously with immunization of said animal with the antigen.

31. A commercial package according to claim 23, wherein the antigen is in a form capable of being applied gastro-intestinally after immunization of the animal by the antigen.

32. A commercial package according to claim 23, wherein the antigen is in unit dose form suitable for application in an enteral manner in a quantity of 1 to 500 mg/kg of body weight.

33. A commercial package according to claim 23, wherein the antigen is in unit dose form suitable for immunization in a quantity of 1 to 500 mg/kg of body weight.

34. A commercial package according to claim 23, wherein the antigen is in unit dose form suitable for parenteral administration in a quantity of 1 to 100 mg/kg of body weight.

35. A commercial package according to claim 31, wherein the antigen is whey.