WO2001068149A2 - Method and use for biologically marking animals and animal products - Google Patents

Abstract

The invention relates to a method for biologically marking animals and animal products by supplying at least one defined immunogen to the living animal by oral way, e.g. in the fodder or drinking water of said animal. Changes which are thus caused in the animal are detected and are used for detecting the origin of the animal or the products thereof by means of an allocation key.

Description

 Process and use for the biological labeling of animals, animal products and animal products

The invention relates to a method and a use for the biological identification of animals, animal products and animal products according to the preamble of claim 1 and from other independent patent claims.

Methods of this type are known from DE 196 43 682 C1 and WO99 / 36 775. The methods serve to prove the origin of farm animals, in particular cattle, pigs and the like, and comprise the following steps: a) biological marking of the live farm animal by application of at least one antigen which is harmless to the farm animal and humans, and b) identification of the antigen by the application of the at least one antigen specific antibody formed by an enzyme immunological or immunochemical detection method.

The starting point for the present new invention lies in the ever closer convergence of global trade. The associated opening of borders, for example within Europe, brings alongside the desired ones economic relief also complicates the inspection of goods. The origin of our food is often not clearly understandable for consumers. In the meat trade, consumers have recently been startled by the appearance of beef contaminated with BSE or poultry reared with feed mixed with dioxon.

In the case of large farm animals, such as cattle and pigs, the animals are provided with ear tags according to the current legal regulations for labeling. Since 01.01.1998, EU-wide regulations for labeling and registration have been in effect for cattle, which in Germany are still supported by the Beef Labeling Act. These regulations stipulate the labeling of cattle with two ear tags and the creation of accompanying documents from which the origin of the animal can be derived. This measure only applies as long as the ears are on the animal or carcass and the accompanying documents are not manipulated. Before slaughter, the animals are grouped according to their origin and the carcasses are then stamped with origin-specific stamps. After dismantling, carcasses can be mixed and no longer clearly assigned.

In the case of smaller farm animals, e.g. Poultry, individual labeling is no longer feasible for cost reasons. This naturally facilitates the manipulation of the accompanying documents, since the animals or carcasses can no longer be clearly assigned to the papers. As a rule, there is no reliable proof of origin for small farm animals.

According to the current state of science, it would be possible to clearly identify and identify farm animals using two biological methods: - The first possible procedure is based on individual-specific sequence variants of the deoxyribonucleic acid (DNA) of each animal. A piece of tissue is removed from the animals, for example when attaching the ear tag, which serves as a DNA marker. A genetic fingerprint of this tissue sample can be created via sequence variants of repetitive elements (microsatellite marking) or via single nucleotide polymorphisms (SNP pattern) of the DNA and stored in a database. By comparing it to the database, each piece of beef can be individually assigned due to its genetic uniqueness. DNA labeling requires expensive analysis of the genome of each animal and consistent tracking of the CV of each individual cattle in the database. In addition to the costs of sampling, there are definitely costs for genome mapping and the recording and tracking of the résumé in the database, even if no carcass control is necessary after slaughter.

- The second possible procedure is based on the principle of immunological labeling, which is comparable to conventional vaccinations. The livestock are given synthetic substances against which the animals form antibodies that can be detected after slaughtering and cutting, for example in meat drip juice. By combining different marking substances, an individual or origin-specific marking should be possible. This method has the advantage over DNA labeling that not every animal in a herd has to be labeled individually. In other words, an inventory marker is possible in which each individual in the inventory has the same marker. This reduces the amount of data required enormously, since a stock can always be given the same marking as it grows. The disadvantage of this form of immunological labeling is that each animal must be vaccinated individually by an appropriately trained specialist. This vaccination can be given by injections or via the nasal mucous membrane using a nasal spray or via an implant. This is time-consuming and costly and is currently only acceptable for cattle, since the profit margin for a slaughter animal does not allow any further costs during rearing for smaller animals, even pigs.

If one compares the DNA labeling with the immunological labeling, the cost aspect clearly speaks for the immunological approach.

The object of the invention is to improve the principle of immunological labeling in such a way that labeling individuals and populations of all animal species capable of producing antibodies becomes technically and economically practicable. The invention is intended to make it possible to determine the origin of animal species that cannot be detected with the prior art.

This object is achieved according to the invention by a method for the unique biological identification of animals, animal products and animal products in that a) that the living animal at least one defined immunogen, which is harmless to the animal, humans and the environment, orally b) that changes produced by the immunogen in the animal, such as. B. against the immunogen directed receptor structures or against the immunogen formed specific antibodies are detected with a responsive to these changes specific detection method, and c) that the location of the origin of the animal, the animal product or the animal product is clearly demonstrated using the detected changes via a predetermined allocation key.

Furthermore, the above-mentioned object is achieved by a method for the unique biological labeling of animals, animal products and animal products in that at least one defined immunogen which is harmless to the animal, humans and the environment is administered to the living animal by oral route, and that an immunogen is used as an immunogen which does not occur in the food chain and in the environment at the animal's place of life.

Furthermore, the stated object is achieved according to the invention by the use of at least one immunogen which is harmless to humans and animals for the biological identification of animals, animal products and animal products by oral route on the living animal and by the use for immunological detection of the animal by the immunogen generated changes and for detection of the origin of the animal, the animal product or the animal product according to at least one of claims 1-17.

The invention also includes a method for determining the origin of animals to which at least one defined immunogen has been administered, and of animal products and animal products from such animals, characterized in that changes which have been produced in the animal by the at least one immunogen supplied to it, such as B. against the immunogen directed receptor structures or against the immunogen formed specific antibodies, are detected with a responsive to these changes specific detection method, and that the origin of the animal, the animal product, or the animal product is clearly demonstrated using the detected changes via a predetermined allocation key

According to a preferred embodiment, the invention for solving the above-mentioned object also relates to a method for determining the origin of animals to which at least one defined immunogen has been administered and of animal products and animal products from such animals, characterized in that changes caused by the animal that at least one immunogen which has been administered orally to it, such as, for example, receptor structures or specific antibodies formed against the immunogen, is detected using a specific detection method which reacts to these changes, and that using the detected changes via a predetermined assignment key the origin of the animal, of the animal product or the animal product is clearly demonstrated

Further features of the invention are contained in the subclaims

In particular, a hapten coupled to a suitable carrier, an antigen, a DNA vaccine, a modified protein, a DNA expression vector, a synthetic vector, a hybrid vaccine, a combination of different immunogens, a mucosal adjuvant can be added Immunogen can be presented together with a stopper that counteracts immunogen degradation

According to preferred embodiments of the invention, the at least one immunogen is immunologically added to the feed and / or the drinking water added marking animal so that the animal ingests the at least one immunogen orally in this way.

According to a preferred embodiment of the invention, the detection of the changes produced by the at least one immunogen in the animal is carried out by detecting these changes in the living or dead animal, in parts of the dead animal, in body fluids, in animal excretions and / or in animal products of the immunologically labeled animal , Parts of the dead animals can be, for example, muscle meat, bones and bone marrow. Body fluids of live and / or dead animals are especially their blood, saliva and meat juice squeezed out of their flesh. Products from such animals are in particular milk and eggs.

In the context of the invention, “immunogen” is understood to mean an agent which stimulates the animal's immune system, which takes the immunogen orally, to a reaction, for example to the formation of antibodies.

All substances which do not occur in the food chain and in the environment at the animal's place of life and are therefore suitable for associating animals labeled with such a place of origin or place of life can be used as the immunogen. This place of origin or place of life can be the place of birth of the animal, but it can also be a later place of life of the animal, for example where it is believed to be fattening. The animal can be marked both at the place of origin and at the fattening site (where it is fattened) by ingestion of one or more immunogens. In this case, each of these locations can later be verified when the animal changes location or in animal products or animal products. In this way, a large number of individual animals can be marked differently. However, it is also possible to mark entire herds or certain animal species with the same marking.

Vaccines which are usually used for therapeutic treatment can also be used as the immunogen, provided that such vaccines are normally not used for the animals to be labeled by an immunogen. For example, a vaccine could be used to label cattle as an immunogen, which is commonly used to vaccinate foxes against rabies, since such a vaccine is not normally used to vaccinate cattle. The rabies vaccine for foxes is an immunogen that is not found "in the food chain and in the environment at the place of life", for example of cattle, pigs and poultry. It is thus an example of what is meant in the present description by the term "in the food chain and in the environment at the animal's place of life", to which the immunogen for origin marking is supplied by oral route.

A great advantage of the invention is that not only those substances which have a therapeutic effect are suitable as immunogens, but also "waste products" from research which cause a reaction in the immune system of animals, but no therapeutic effect, for example do not create protection against a particular disease.

According to the invention, a single immunogen or several immunogens of different types can be used, either in combination or one after the other to mark the animals at different ages or at different places of life.

Oral immunization can significantly reduce the costs of the labeling process if suitable immunogens, individually or in defined combinations, are added to the feed or drinking water. Labeling is inextricably linked to the labeled animal and the products derived from it by this method, as in the injection of the immunogen. Location, date and an animal definition etc. are stored in a database or in some other way, so that this data can be used later to identify the animal's origin or species or to identify special living conditions of the marked animals, if later on the living or dead animal , on whose products or products the "changes" of the animal "generated" by the specific immunogen or the specific immunogens are detected. Through an "assignment key", which e.g. a comparison table e.g. can be in an electronic database or on another data carrier, the "generated changes" can be clearly assigned to a defined immunogen and from this to the marked animal or the marked animal species and the latter can be recognized.

The cost reduction allows not only to mark cattle stocks, but also smaller animal species. The marking is not limited to mammals (pigs, goats, sheep, rabbits, etc.), but the invention is applicable to all living beings that are capable of producing antibodies. At this point it should be mentioned explicitly e.g. B. the possibility of marking poultry (chickens, turkeys, geese, ducks etc.) and fish stocks (eg salmon, trout, carp etc.). In the case of immunological labeling of poultry, the ones formed Antibodies, for example, can also be detected in the eggs of the marked animals. By marking laying hens, it is also possible to prove the origin of eggs. The marking of cows makes it possible to prove the origin of milk, since the reaction to immunogens can also be detected in milk. This form of immune labeling can contribute to the protection of species in exotic animal breeding, such as turtles, in order to distinguish, for example, meat products from labeled breeding animals from unmarked slaughtered wild animals.

For the immunization, poorly digestible antigens, preferably simple molecules and conjugated vaccines, or DNA vaccines are preferably used, which do not occur in the food chain or generally in the environment at the place of origin of the animals and which are also not used as therapeutic vaccines for the animal to be identified. The design feature of the immunogens is to overcome all physiological and immunological barriers after oral intake in order to form the specific antibodies, which can be detected using an enzyme-immuno or immunochemical method. Modified proteins or haptens coupled to suitable carriers are used as immunogens, which are harmless to the animal and harmless to human consumption. In addition to peptides, synthetic or natural polymers, for example naturally non-occurring or non-locally occurring enantiomers of natural products, are also suitable as haptens. Recombinant or attenuated bacteria and viruses in connection with mucosal adjuvants or protease-resistant bacterial toxins can be used as the haptic carrier. DNA expression vectors and synthetic vectors, for example proteoliposomes and lipoproteids, are also suitable as immunogens. All these foreign and harmless molecules for the animal organism enable according to the invention individually or in Combination a unique biological labeling of animals, animal products and animal products.

The use of a mixture of different immunogens is advantageous in order to increase the number of labeling options, and the antigenic mixture can also be administered on a single hybrid vaccine. Certain immunogens can stand for the identification of a region or a certain animal species. The further combination of immunogens allows the pooling of animal populations.

An advantage of oral immunization by ingestion of the immunogens with the feed or drinking water is that the antibodies formed, although desirable, but not necessarily, have to remain permanently in the body of the immunized animal, since booster immunizations can be repeated periodically at low cost. This ensures that a uniformly high titer of immunogen-specific antibodies is present after the immune response has responded. The age of the animals to be immunized plays a subordinate role due to a periodic refreshment. The species-specific immunization cycle is more important. The species-specific and / or posture-specific cycle length ensures that each animal comes into contact with the immunogen or immunogens at least once in the most sensitive period of its type for immunization and can form the corresponding antibodies. These periodic immunizations also ensure that long-lived breeding animals or their products can be clearly identified and identified.

The identification of the antibodies or receptor structures specific for the labeling, for example T-lymphocytes, can be carried out using various immunological detection methods. For example, The following highly sensitive immunoassays: Enzyme Immuno Assay (EIA), Enzyme Linked Immunosorbent Assay (ELISA), Fluorescent Immuno Assay (FIA), Radio Immuno Assay (RIA), Luminescent Immuno Assay (LIA) etc. Simpler but generally less sensitive are immunochemical test platforms such as lateral flow and flow-through assays, which can also be used for antibody detection. The uniformly high antibody titer against the immunogens used, as is achieved with cyclic immunizations, is of great advantage for the detection of the label.

The identification of the changes produced by the immunogen in the animal can also be carried out by molecular biological detection methods, e.g. B. Southern blot, Western blot, PCR (polymerase chain reaction), or biochemical detection methods, e.g. B. Use of enzyme-catalyzed reactions.

The number of possible combinations for the unique labeling of individual animals, individual animal species or an animal population depends on the number of immunogens used for the immunization and the total number of immunogens available for the combination.

Based on an available quantity of, for example, 96 different immunogens, which corresponds to the maximum occupancy of a conventional microtiter plate, over 900 are clearly marked for individual animals, individual animal species or an animal population with a combination of 6 immunogens that are added to the feed or drinking water Millions of possible markings. This number is more than sufficient to characterize the global livestock population and gives the necessary scope to not only achieve a primary marking (marking by the young animal producer), but also animals in the secondary branch (marking at different locations of the animal after birth, in a fattening farm, for example) In this case, 48 immunogens can be used for primary labeling. Another 48 immunogens can be used for labeling in secondary masts. For primary and secondary labeling, there are approximately 12 million possible combinations available

If an ELISA system is used as the detection method, in which an immunogen is assigned to each well, a pattern of six immunogens of the primary immunogen class is obtained when the primary labeling is detected. If a secondary mast is detected, another pattern of 6 immunogens of the secondary immunogen class is added to this pattern in addition

The invention thus enables the immunological labeling of animals of any kind. Not only individual animals as individuals, but also various animal species or certain animal populations, in particular livestock populations, are suitable for immunological labeling. Vertebrates, which are capable of forming antibodies, are particularly suitable for labeling. especially mammals, but also birds, lizards, fish and amphibians

The labeling is preferably carried out by adding the at least one immunogen to the animal feed or drinking water, preferably in the form of artificial antigens or combinations of antigens. which are harmless to animals, humans and the environment and which cause the formation in the animal of receptor structures or specific antibodies directed against the immunogen. The receptor structures or antibodies produced in response to an immunogen in the animal are later not only in the live animal, but also in the meat and meat juice, in the saliva and blood of slaughtered animals, as well as in the milk and eggs and other animal products of the labeled Animals present and detectable, in particular detectable by an immunological method.

The invention thus relates to a method and its use for the biological identification of animals, animal products and animal products in that at least one defined immunogen is administered to the living animal by the oral route, for example in its feed or drinking water. Changes that are caused in the animal are detected and used to determine the origin of the animal or its products or the animal products using an assignment key.

The invention also relates to a method for determining the origin of animals to which at least one defined immunogen has been administered and of animal products and animal products from such animals, characterized in that changes in the animal by the at least one of them, preferably by the oral route , supplied immunogen were generated, such as. B. against the immunogen directed receptor structures or against the immunogen formed specific antibodies can be detected with a specific detection method reacting to these changes, and that using the detected changes over a predetermined allocation key the origin of the animal, the animal product, or the animal product clearly is proven. The detection of the changes produced by the immunogen in the animal is preferably carried out by a molecular biological or biochemical or immunological method or a molecular biological or biochemical or immunological rapid test method.

Claims

claims 1. A method for the biological identification of animals, animal products and animal products, characterized in that a) that at least one defined immunogen, which is harmless to the animal, humans and the environment, is orally administered to the living animal, b) that changes caused by the immunogen in the animal, such as. B. against the immunogen directed receptor structures or against the immunogen formed specific antibodies, are detected with a reaction to these changes specific detection method, and c) that using the detected changes via a predetermined allocation key, the location of the origin of the animal, the animal product or the animal product is clearly demonstrated. 2. A method for the biological identification of animals, animal products and animal products, characterized in that the living animal is given at least one defined immunogen, which is harmless to the animal, humans and the environment, by oral route, and that an immunogen is an immunogen is used, which does not occur in the food chain and in the environment at the animal's place of life. 3. The method according to any one of the preceding claims, characterized in that an immunogen is used which is usually not used as a therapeutic vaccine for this animal at the place of life of the animal to be labeled. 4. The method according to any one of the preceding claims, characterized in that at least one hapten coupled to a suitable carrier is used as the immunogen. 5. The method according to any one of the preceding claims, characterized in that an antigen is used as the immunogen. 6. The method according to any one of the preceding claims, characterized in that a DNA vaccine is used as an immunogen. 7. The method according to any one of the preceding claims, characterized in that a modified protein is used as the immunogen. 8. The method according to any one of the preceding claims, characterized in that a DNA expression vector is used as an immunogen. 9. The method according to any one of the preceding claims, characterized in that a synthetic vector is used as the immunogen. 10. The method according to any one of the preceding claims, characterized in that a hybrid vaccine is used as an immunogen. 11. The method according to any one of the preceding claims, characterized in that a defined combination of at least two different immunogens is used. 12. The method according to any one of the preceding claims, characterized in that at least one mucosal adjuvant is admixed to the immunogen, which is harmless to the animal and human consumption. 13. The method according to any one of the preceding claims, characterized in that the immunogen is presented together with a substance which counteracts the immunogen breakdown. 14. The method according to any one of the preceding claims, characterized in that the immunogen is added to the feed and is ingested in this way by the animal orally. 15. The method according to any one of the preceding claims, characterized in that the immunogen is added to the drinking water of the animals and is ingested orally by the animal in this way. 16. The method according to any one of the preceding claims, characterized in that the detection of the changes in the animal produced by the immunogen in the living or dead animal, in parts of the dead animal, in body fluids, in animal waste and / or in animal products such as, in particular, milk and Eggs. 17. The method according to any one of the preceding claims, characterized in that the detection of the changes generated by the immunogen in the animal is carried out by a molecular biological or biochemical or immunological method or a molecular biological or biochemical or immunological rapid test method. 18. Use of at least one immunogen which is harmless to humans and animals for the biological identification of animals, animal products and animal products by oral route on the living animal and for the immunological detection of the changes produced by the immunogen in the animal and for the detection of the origin of the animal, the animal product or the animal product according to at least a d _er A _Λ nsprü ..ch _.e "1-" 1.7. 20 19. A method for determining the origin of animals to which at least one defined immunogen has been administered, and of animal products and animal products from such animals, characterized in that changes which were produced in the animal by the at least one immunogen supplied to it, such as, for. B. against the immunogen directed receptor structures or specific antibodies formed against the immunogen, can be detected with a specific detection method reacting to these changes, and that using the detected changes over a predetermined allocation key the origin of the animal, the animal product, or the animal product clearly is proven. 20. A method for determining the origin of animals to which at least one defined immunogen has been administered and of animal products and animal products from such animals, characterized in that changes which were produced in the animal by the at least one immunogen which is supplied to it by oral route, such as B. against the immunogen directed receptor structures or against the immunogen formed specific antibodies can be detected with a specific detection method reacting to these changes, and that using the detected changes over a predetermined allocation key the origin of the animal, the animal product, or the animal product clearly is proven. 1. The method according to claim 19 or 20, characterized in that the detection of the changes produced by the immunogen in the animal is carried out by a molecular biological or biochemical or immunological method or a molecular biological or biochemical or immunological rapid test method.