DK172775B1 - Animal feed additives containing a flavanoid-containing mimosa extract and animal feed containing the same and method t - Google Patents

Description

in DK 172775 B1

The present invention relates to an animal feed additive comprising a flavanoid-containing extract derived from a plant belonging to the family Mimosaceae. and an animal feed containing such an additive. The additive can be given directly to animals in need, but is especially intended for admixture in a feed or beverage intended for the animal (s) concerned. The additive and feed or beverages containing it are capable of controlling and / or preventing diarrhea and other gastrointestinal disorders or disorders in animals, promoting the growth of and / or enhancing feed utilization in animals fed with the additive or feed. or drink.

10 The additive and feed additive are intended especially for agricultural production animals, such as pigs, cattle, sheep, goats, horses and chickens and other poultry and their offspring, such as pigs and piglets, calves, lamb, kid, foal and chickens, etc. but is also useful for pets, such as dogs and cats and their offspring.

Background of the Invention

Over the past few decades, there has been a greater and greater rationalization, specialization and intensification in agricultural livestock production, so that at each farm today there are usually only very few, yes, many times only one person, that the demand for high productivity and short production time is extremely large and in most 20 cases only a single animal species is produced, e.g. slaughter pigs, broilers or beef cattle, or the products of this species, e.g. piglets from pig sows, milk from dairy cows or eggs from laying hens. On the other hand, the species of animal in question is kept in a very large number from hundreds to many thousands of individuals on the individual farm 25

These conditions mean, firstly, that virtually all care and feeding of the animals must be automated and can be performed mechanically, secondly, that the animals are constantly subjected to a great deal of pressure due to the many animals of the same species on a single holding. and thirdly, that the physiological systems of the animals, i.a. the digestive system, due to the requirement 30 for high productivity and short production time, is constantly subjected to such great pressure that even less extra loads can cause the systems to collapse, which for the digestive system e.g. can result in severe diarrhea, which either causes the animals to become delusional or possibly dies after a shorter or longer period, which in both cases results in loss of production for the farmer.

Furthermore, the requirement of high productivity / shorter production time is also expressed by the fact that the offspring of the animals, e.g. weaners and baby calves, weaned, ie. are removed from the maternal animal and thus from their previous natural nutrition, the breast milk, at an earlier stage of their life than was previously the case, which for the animal cubs was a physiologically more appropriate time in their development and maturation to the independent adolescent level. Thus, piglets today are usually weaned at the age of just 4-5 10 weeks after giving birth, compared to the previous 8-9 weeks.

The early weaning of, for example, the pigs and the transition to exclusively other nutrition than the sow's milk inevitably leads to an immediate or after a brief onset of chronic diarrhea in almost all piglets, and without treatment these 15 pigs will not only be delayed for a long time thereafter but in many cases simply die.

The above relationship has been known as. an inevitable consequence of the evolution of production in the livestock farms, and when this has nevertheless been possible, it is only by virtue of a simultaneous increasingly widespread use of antibiotics of various kinds to the 20 animals, whether administered as parenteral injections in the animals or, what more commonly, mixed with animal feed. In the latter case, the antibiotic additives in question are commonly referred to as "growth promoters".

The antibiotics used for the animals, e.g. however, growth promoters tend to cause resistance in the bacteria and other microorganisms present in the animals, and it has therefore been necessary to constantly develop and use new antibiotics for the animals. However, some of the antibiotics used in animals are the same or chemically so closely related to the antibiotics used to control diseases caused by bacteria and other microbes in humans, that in the public opinion, especially in recent years, there have been a growing concern that the resistance that occurs in the microorganisms in the animals should be transmitted to the human disease-causing microorganisms and that these could not be combated with the human antibiotics available.

Against this background, there is therefore an ever-increasing demand to limit the use of 5 antibiotics of all kinds to animals as much as possible and to completely avoid, indeed, directly ban the use of antibiotics of all kinds as "growth promoters" in the animal feed completely.

From the above statement on the conditions of production in today's livestock farms, it will be understood that such a ban on growth promoters will of course result in catastrophic conditions for both animals, production results and livestock farms, unless previously provided alternative treatment methods and agents which are not based on substances. antibiotic nature, and therefore a great deal of research and research is currently underway for such methods and means, both for brand new and among known and previously used, which for some reason have become out of use during or 15 before "period of antibiotic growth promoters".

Prior Art WO Publication No. 96/03150 thus describes compositions of hemicellulose fibers and polyphenols derived from larch trees fLarix occidentalix NuttallV 20 The exemplified hemicellulose fibers and polyphenols thus consist of the water-soluble arabinogalactan and taxi poles, respectively. These compositions are said to have a growth-promoting effect on beneficial bacteria such as Bifidobacterium thermophilum. and an inhibitory effect on harmful, pathogenic and decay bacteria such as Clostridium ssp .. Bacteroidaceae and Candida ssp .. in the gastrointestinal tract of humans and animals and thereby fight gastrointestinal disorders such as diarrhea. It is mentioned that flavanoids and tannins fall within the group of polyphenols which could be used in the compositions of this invention, but this has not been exemplified or otherwise demonstrated. The ratio of fibers to polyphenols in the compositions is said to be between 20 and 3, and such compositions are said to be capable of being added to animal feed to the extent of 0.1-5% by weight, preferably 0.1-2% by weight, based on the weight of the animal feed. . However, in the document only experiments with pure arabinogalactan or pure compositions of arabinogalactan and polyphenols extracted from larch wood have been described in connection with testing the effect on various Bifido bacterial strains grown in test tubes containing an amplified clostridial agar soup, but no forms of experiments. on feeding animals with a feed containing arabinogalactan polyphenol compositions is disclosed. It is therefore not known whether the composition 5 or a feed containing it will work in practice, or whether e.g. There are problems in getting the animals to voluntarily eat such feed. - It is further stated that the compositions may be formulated into tablets, pills, capsules, lozenges and syrups, and that these dispensing forms may contain sweeteners such as sucrose or saccharin and flavors such as peppermint, methyl salicylate or orange flavor. These forms of dispensing are normal to humans but not to animals, and there is then no indication that they should be intended for admixture or in any way intended for animal feed.

Another known agent that has been used in the livestock farms for the control and / or prevention of diarrhea conditions in animals, especially in infants and weaners, infants and other animal infants, before the discovery of antibiotics and especially before the emergence of growth promoters, is a flavanoid-containing extract. extracted from a plant belonging to the Mimosaceae family. This extract was previously sold under the trade name "Calf and Pig Relief" and later under the name Diarex® in the form of either a mixture containing, in addition to water 20 and a small amount of preservative, approx. 50% by weight of the flavanoid-containing plant extract, or in the form of a powder which, apart from a minor amount of anti-caking agent, consisted of the pure flavanoid-containing plant extract. The agents were administered directly to the animals as they were available at the recommended doses (2 - 30 ml or g per animal per day) or mixed with the animals' feed or drink.

25

However, the mimosaceae extract has an odor and / or taste repellent to the animals, and therefore the animals will generally under no circumstances voluntarily ingest it, whether as a mixture, powder or in their feed or drink. In the past, therefore, it was necessary that each animal during administration must be kept by one or more persons, while another (or third) person with a syringe or feed or powder can sprayed the mixture / drink or stuffed the powder or the mixture or powder added respectively. feed directly into the animal's esophagus. In small animals, such as infants and calves, it was an affordable, albeit cumbersome and time-consuming task to treat each animal up to several times daily in this way, but for larger animals it was due to the physical size of the animal and Strength almost impossible unless special devices were used to hold and lock the animal in an immobile position.

The treatment or prevention of diarrheal conditions in the animals with the Mimosaceae extract in the above manner was in itself effective and inexpensive, but because of the cumbersome and time-consuming mode of administration, it was practically completely out of use, since Antibiotics and growth promoters came on the market at affordable prices, as these slugs had no odor and / or taste to the animals and could therefore be mixed directly into the animals' usual feed, which the animals would then voluntarily eat.

15 In the present situation, however, where the use of antibiotics as growth promoters and anti-diarrheal and preventative agents for production animals probably must be completely abandoned in the near future in an ever-increasing number of countries, the inventors of the present invention have come to the idea that it would be desirable, if possible, to change the character of the Mimosaceae extract so much and / or develop a method of administration thereof which was not beset with the difficulties mentioned above, as the animals would not voluntarily drink or eat the extract-containing mixture / drink and powder / feed, respectively, since the Mimosaceae extract contains no antibiotic substances and is not absorbed from the gastrointestinal tract in the animal organism at all.

25 Initially, it was thus attempted to incorporate the Mimosaceae extract into the usual feed of the animals in such a low concentration that the animals were not repelled by smell and / or taste and therefore would eat the feed voluntarily, but due to the very fine and well-developed sense of smell and taste, it was found that the low concentration required for this was so small that a sufficient therapeutic and / or prophylactic effect could not be achieved by the amount of feed voluntarily ingested by the animals.

6 DK 172775 B1

It was then attempted to encapsulate the Mimosaceae extract powder as coated (coated) particles or granules which were then blended into the animal's feed, but again due to the animals' extremely delicate sense of smell and / or taste, this method also did not give a satisfactory result of the same reasons as stated above. Furthermore, the coating process means that the product became too expensive to be used as a commonly used feed additive in practical animal breeding and production.

Description of the Invention

It is therefore very surprising that, with the present invention, it has now nevertheless proved possible to provide the Mimosaceae extract in a special and novel additive form which allows the animals, such as, for example, weaning pigs and calves, to voluntarily take the additive form, either directly as it is, or in the usual feed of the animals or drink in sufficient quantities to achieve both therapeutic and prophylactic effect, particularly with regard to the control and prevention of diarrhea conditions in the animals.

This is achieved with the feed additive according to the invention, which comprises a flavanoid-containing extract derived from a plant belonging to the family Mimosaceae. which feed additive is characterized in that it further comprises an odor and / or taste masking substance 20 or a mixture of such substances which mask the odor and / or taste of the flavanoid extract, and optionally one or more carriers for the odor and / or flavor mask or the mixture thereof.

In addition to the above ingredients, the additive may also contain other ingredients, such as actual fillers and / or extenders, dyes, consistency conditioners, essential minerals, vitamins, amino acids, etc., which do not repel the animals or reduce their appetite or appetite.

Preferred embodiments of the additive according to the invention are set out in the dependent claims 1-7.

For the invention and the embodiments thereof, the following observations should be made: 7 DK 172775 B1

The feed additive according to the invention will preferably be in dry powder or minigranulate form but may also be available in moist, paste or liquid form.

Regardless of the physical state, the additive can generally be prepared by simple, thorough mixing of the ingredients. Thus, a powdery additive can be prepared by dry mixing the ingredients, but it can also be prepared by wet mixing the ingredients followed by drying, grinding and possibly sieving. A minigranulate can be prepared in the same way, however, initially it may be necessary to add sufficient liquid of suitable nature (eg water) to achieve sufficient cohesion between the particles of the ingredients.

10

By an odor and taste masking agent is meant any non-harmful substance capable of sufficient concentration to mask the unpleasant and repulsive odor and / or taste of the Mimosaceae extract to the animals that the animals will voluntarily eat enough of the additive or a feed / beverage in which the additive is mixed to allow the animals to ingest a therapeutic and / or prophylactic amount of the Mimosaceae extract. The odor and / or taste masking agent can exert its masking effect in various ways. For example, it may: 1) have in itself such an appealing and appealing odor and / or taste that overpowers or blends with the smell and / or taste of the Mimosaceae extract, so that the latter is neither perceived nor works 2) exert a blocking effect on the receptors in the olfactory and / or taste cells of the animals' nose and mouth that register the Mimosaceae extract as odorous and / or taste unpleasant, 3) bind physically / chemically to the substances or molecular groups of substances in the Mimosaceae extract which cause the odor and / or taste to be repulsive to the animals, thereby blocking their odor and taste effect, provided that the Mimosaceae extract does not thereby lose its therapeutic and prophylactic effect, or 4) any combination of the above properties or effects.

30

Examples of odor and / or taste masking agents which exert their masking effect in one or more of the above-mentioned ways are set forth in claims 6 and 7. Particularly preferred is an additive containing 5-80% by weight of flavanoid containing. Mimosaceae extract and 95-20% by weight of biscuit flour, preferably 10-50% by weight of extract and 90-50% by weight of biscuit flour, more preferably 15-30% by weight of extract and 85-70% by weight of biscuit flour, in particular approx. 20% by weight of extract and approx. 80% by weight of cookie flour, based on the total weight of the additive.

5

The possible incorporation of one or more carriers for the odor and / or taste masking substance into the feed additive according to the invention will depend on the specific substance used and the purpose of its incorporation will accordingly be contingent upon it. For some very strongly smelling and / or tasting masking agents, a carrier will be necessary to simply obtain a sufficient dilution of the former to be able to obtain a suitable dosage and homogeneous mixing with the Mimosaceae extract. For other masking agents, a carrier will be required to be able to retain the masking substance intact in the additive mixture, e.g. to counteract the evaporation, excretion, degradation, or other harmful conversion of the substance during storage of the additive for a shorter or especially longer period. In other cases, again, the function of the b-honour may be to provide a sufficiently close association of the masking substance with the smelling or tasting substances of the Mimosaceae extract to achieve the desired masking effect.

Any carrier which is capable of providing the above effects and which has no detrimental effect on the therapeutic and prophylactic effect of the Mimosaceae extract can be used as a carrier, while of course it should not in itself have any odor and / or taste that will repel the animals. The carrier can be of organic or inorganic, digestible or indigestible nature. Examples of carriers include carbohydrates such as starch particles, cellulose fibers and mono-, di- and oligosaccharides, protein substances, e.g. soy protein, fats and oils, as well as salts such as calcium phosphate and sodium chloride, as well as mixtures of such substances. - Examples of substances which are not commonly used as carriers may be mentioned ferrous compounds and substances of various kinds, since the iron in these compounds or substances will in many cases block the effect of the therapeutic and prophylactic effects of Mimosaceae extract.

9 DK 172775 B1

That the additive is in powder form means that the largest diameter of the additive chair particles does not exceed approx. 0.2 mm. The fact that the additive is in micronutrient form means that the main amount, e.g. at least 80% by weight, preferably at least 90% by weight, more preferably at least 95% by weight, and most preferably at least 98% by weight, of the particles having a maximum diameter which is in the range of 0.2 to 2.0 mm.

As for the flavanoid-containing extract itself, it is extracted as mentioned by a plant belonging to the family Mimosaceae. These plants are tree or shrub-like growths, which grows in southern Africa, where they are grown in various places in plantations that are harvested, 10 when the bark of the plants is considered ripe for the extraction of the flavanoid-containing compounds therein. That is, in the case of woody growths, when the plants are usually at least approx. 9-10 years old. The harvest usually takes place during the growing season of the plants, ie. in September-March, when the bark has a moisture content of approx. 50%. The plants are barked and the bark is transported, possibly after prior drying to bark sticks, to extraction plants in the country or abroad where it is cut into shavings subjected to an autoclave extraction process with water at superatmospheric pressure and a temperature above 100 ° C. or an extraction by percolation with lower temperature water after rupture of the bark parenchyma cells by autoclaving. After concentrating the extract liquid, it is spray-dried in a countercurrent spray-drier to produce a reddish, yellowish or greenish-brown powder having a moisture content of approx. 6%.

The powder, which consists of hollow spherical particles, has a basis weight of from 370 to 500 kg / m3, preferably 370 to 430 kg / m3, is more easily hygroscopic and is soluble in e.g. water and alcohol. A 13% solution of the powder in water has a pH of 4.6, a smaller buffer range 25 between Ph 2.8 and 5.8, and a larger buffer range at Ph 9.5. Aqueous and alcoholic solutions are stable for prolonged storage in the dark, but exposure to sunlight will cause redness. The powder is stable at dry and dark at room temperature for at least 3 years. Exposure to sunlight will result in red staining of the surface layer, and upon heating, initial charring will occur at over approx. 230 ° C. The content of polyhydroxy flavanoids in the powder will depend on the particular Mimosaceae variety. of which the extract powder is obtained, but will generally be in the range of 65-75% by weight, in particular about 67-72% by weight, in particular approx. 70% by weight. By mixing two productions, equalization can take place in the 67-72% range.

An alcoholic (99.8% ethanol) extract of the powder in a 0.25% (w / v) solution by adding 0.25 ml of 1% FeCl 3 solution to 10 ml of extract solution will give a clear blue solution without precipitate. Presence of tannic acid will give a blue precipitate after centrifugation and should not occur.

Thin layer chromatography on silica gel plates of the alcoholic (99.8% ethanol) extract of the powder will be developed with butanol / acetic acid / water (dist.) In the 4: 1: 5 mixture ratio to give

Pipe values that depend on the particular Mimosaceae variety. from which the extract powder is obtained, and possibly also from the extraction conditions. Extract powder from Acacia me-amsii De Willd. which grows like trees in South Africa, and is obtained by autoclave extraction with water at ca. 110 ° C for approx. 6 hours, thus has an Rf of approx. 0.70, while ex-15 funnel powder from Herba Acacia Catechu Willd. black Heleati. Am., Which grows like shrubs or herbs in Southwest Africa (Namibia), and which is obtained by water percolation for 48 hours after autoclave bursting of the parenchymal cells, has an Rf of approx. 0.35. These values must be seen in relation to tannic acid having an Rf of 0.9 under the same conditions.

Other physical and chemical characteristics of Mimosaceae extract powders can be found in the specialty literature.

As mentioned, 65-75% by weight of the Mimosaceae extract powder is made up of polyhydroxy flavonoids, while the remainder is a complex mixture of carbohydrates, gums, sugars, amino acids and other minor constituents, of which a small amount of insoluble matter. The part constituted by the polyhydroxy flavanoids consists of condensed flavanoids and their monomeric precursors. Thus, the polyhydroxy flavanoids comprise a complex mixture of a number of different single components, the basic unit of which is the typical carbon-15 flavanoid skeleton, and their molecular weight varies from ca. 300 for the 30 nominal precursors to ca. 3000 for the largest polymers with an average molecular weight of approx. 1200-1300. Accordingly, the average Mimosaceae flavanoid molecule consists of 4-5 monomeric flavanoid molecules. They are bonded to each other with carbon-DK 172775 B1 π carbon bonds. The flavanoid precursors are typically constituted by flavan-3,4-diols (leucoanthocyanidines) and flavan-3-ols (catechins), see Schemes 1 and 2 below.

When decomposing the flavanoid molecules, the heterocyclic C-ring 5 is typically disrupted to produce resorcinol, phloroglucinol and protocatechinic acid, gallic acid and smaller amounts of resorcinol, phloroglucinol and catechol, pyrogallol, respectively, see Schemes 3 and 4.

The polymeric flavanoid molecules are predominantly constituted by flavan-3,4-diols condensed Γ0 with flavan-3-ols. The structures of the di- and triflavanoides are shown in Figures 5 and 6.

The polyflavanoids have a strong affinity for collagen, which forms the basis for their use in the traditional vegetable tanning methods. The true affinity of the flavanoid compounds for proteinaceous material first becomes clear from the level of the triflavanoid level, as can be demonstrated by the triflavanoids precipitating gelatin from a solution under controlled conditions. Further information on the chemical composition and stereo structures of the Mimosaceae flavanoids and on the content of non-flavanoid compounds of the Mimosaceae extracts can be found in the special literature.

The Mimosaceae extracts themselves have no direct bactericidal or microbiocidal effect, i.e. they have no direct killing effect on bacteria and other microorganisms. The effect of the extracts on the microorganism flora (bacteria and other microorganisms) in the gastrointestinal tract of animals and humans is believed to be due to the precipitation and binding of proteins from the outermost surface of the intestinal epithelium and into the gut contents and on the surfaces of the microorganisms. Hereby, bacteria and other microorganisms, and in particular their toxins, are bound and wrapped, so to speak, in a protein flavanoid envelope or network, thereby preventing or significantly reducing their deleterious effect on the intestinal wall and greatly reducing their number in the intestine by the intestinal peristaltic ejection of the indigestible protein. -favanoid casing or -30 complex. Also, since the microorganisms are not directly killed, their toxins are not released to a large extent, as is the case with the use of large doses of traditional antibiotics that can lead to toxin shock and the eventual death of the treated individual. Furthermore, in the flavanoid treatment, non-resistant bacterial strains emerge during the flavanoid treatment, as the control is based on a general physico-chemical entrapment and binding of the microorganisms which are then excreted with faeces, rather than on a specific chemical / biochemical intervention in the life of the organisms. - and / or reproductive processes, in which survivors of 5 organisms will be able to develop countermeasures to, and thus generation by generation, become increasingly resistant to the antibiotic's mode of action.

However, after single-dose administration of the Mimosaceae extract, in some cases, there may still be such a large number of bacteria / microorganisms remaining in the gastrointestinal tract that they can be rapidly propagated, thereby continuing or restarting a diarrhea state in the treated individual. Therefore, the mimosaceae extract should generally be administered at least twice daily for 2-4 days and possibly longer to obtain optimal results.

The active ingredients of the Mimosaceae extract are not directly digestible, and neither these nor their degradation products have been detectable in the blood or milk of individuals who have been orally administered the extract. Therefore, oral administration of the extract in an individual results in a local effect only in the gastrointestinal tract, which is why systemic side effects are unlikely to exist and have not been reported. Also, there will be no need for "retention times" for meat or milk from animals treated with Mimosaceae extract.

As the Mimosaceae extract alone has effect in the gastrointestinal tract by physically decimating the amount of microorganisms [such as: A) anaerobic gram-negative rods (bacteroides), cooks (pepto-streptococci and peptococci), clostridia and lactic acid bacteria; B) aerobic (about 1% of total) enterobacteriaceae, streptococcus species and bacillus species; and C) miscellaneous fungi and the like, counteract the tendency to overgrowth of one or a few species, often seen after the use of antibiotics.

In addition to an animal feed additive comprising the above-described flavanoid-containing Mimosaceae extract, the invention also relates to an animal feed or beverage which is characterized in that it comprises a feed intended for that animal or beverage and beverage. containing additives according to the invention, preferably in such weight ratios that the pre-mixed feed / beverage, respectively, comprises 0.05-10% by weight of the flavanoid-containing extract, more preferably 0.1-5% by weight, most preferably 0.5 - 2% by weight, especially approx.

1% by weight, based on the weight of the total feed or drink mix.

5

The additive of the invention may be used to treat and / or prevent diarrhea or other gastrointestinal disorder or disorder in an animal, to promote the growth of an animal and / or to increase feed utilization in an animal, the process of which is provided by the animal. a feed or beverage according to the invention.

10

The additive according to the invention can be used to control Salmonella typhimurium DT 104 in the gastrointestinal tract of animals, the animal being given an additive, a feed or a beverage according to the invention.

Finally, the invention relates to a process for the preparation of a feed or beverage for controlling Salmonella typhimurium DT 104 in the gastrointestinal tract of animals, the method being characterized in that a feed or beverage intended for the animals in question is added to an additive according to the invention. one of claims 1-8, preferably in the weight mixing ratio of claim 9.

20

As with the pure Mimosaceae extract, when treated with the animal feed additive or feed / beverage / drink, after single dose administration of the feed additive or feed additive according to the invention, in such cases, a large number of bacteria / microorganisms left in the gastrointestinal tract that they will be able to rapidly propagate, thereby continuing or restarting a diarrhea state in the treated individual. Therefore, the feed additive or feed containing feed according to the invention should generally be administered at least twice daily for 2-4 days and possibly longer to obtain optimal results. However, the great advantage of using the feed additive and / or feed / drink of the invention is that while the pure Mimosaceae extract must be administered to the animals by forced feeding, the animals will voluntarily and with great desire and eagerness take the additive and / or feed / drink. according to the invention, thereby obtaining the beneficial therapeutic and prophylactic effect without the use of time-consuming and costly labor effort.

Thus, by feeding animals with the additive, feed or beverage of the invention, it will be possible to prevent or control any diarrheal or other gastrointestinal disorder in the animals and to combat or prevent the presence of any unwanted bacteria or other microorganism in the animals. gastrointestinal tract, including e.g. bacteria belonging to the E.Coli families. Salmonella (e.g.

S.choleraesuis and S.tvimimuriumk Clostridium (e.g.

10 Sernulina (e.g., S. hemodvsenteria and S.pilosicoli) and Lawsonia (e.g., L. intracellularis).

The invention will be further elucidated in the form of subsequent exemplary embodiments and test results.

Example

An animal feed additive according to the invention having the following composition was prepared: 20 Component amount by weight% kg

Biscuit flour (biscuit flour) 400.00 80.00 25 Diarex® powder, wife. 100.00 20.00 1 Total 500.00 100.00 30

Diarex® powder, wife. is a Mimosaceae extract powder containing ca. 1% o anti-caking agent.

15 DK 172775 B1

Biscuitsmeal manufactured and sold by Internationale Handelmaatschappij Robi bv,

The Netherlands is specially designed for young animals and consists of a mixture of 70% by weight of ground biscuit and pastry products and 30% by weight bread flour. The product, which is free-flowing, is easily digestible, tasty and has a pleasant vanilla-sugar-like smell.

5

Biscuitsmeal is stated to have the following composition and feeding properties:

Ingredient Analyzed Guaranteed content,% content,% 10 ____

Humidity 10.6 max 11 Crude protein 9.3 min 9 Crude fiber 0.8 max 1.5

Ash 2.2 max 3 15 Sugar + glucose min 18

Starch (pre-cooked) 38

Starch, total 40 Crude fat min 7

Carbohydrates 70.0 67.5 Calcium 0.5

Chlorine (as NaCl) 1.2

feed Safety

Feeding units per 100 kg: 132.8 25 EFOS pigs: 96.9

amino acids

Lysine: 2.2 g / kg

Methionine + cystine: 3.7 g / kg 30

Granulometry 98% max 2 mm

Heavy metals: absent I6 DK 172775 B1

Microbiological quality

Entero bacteria per grams <1000

Salmonella pr. 25 grams absent 5

Biscuitsmeal and Diarex® powder, both available in powder form, were dry mixed by thorough mixing in a Nauti mixer (cone-mixer) for 15 minutes and then available as a dust-free (slightly moist / cohesive, but otherwise essentially free-flowing) powder. minigranulate, which was ready for incorporation into an animal feed or animal drink.

From the prepared feed additive, a dry feed for weaning piglets (age 4-5 weeks) of the following composition was prepared: 15 Ingredients dry matter weight% quantity% kg

Wheat (ground) 86.0 51.50 257.50 20 Biscuitsmeal (biscuit flour) 89.4 10.00 50.00

Fishmeal, ash low 94.0 10.00 50.00

Soy protein, wife. HP 300 92.0 5.00 25.00

Flex Combimilk STD. 97.0 10.00 50.00

Fat, an. pigs, 92/10 99.5 3.50 17.50 25 Feed additive according to. opf. 90.5 5.00 25.00

Flex Norm-Mix 2201 96.7 5.00 25.00 Total 100.00 500.00 30 In the table denotes:

Flex Combi milk STD: Skimmed milk powder with vegetable fat Fat, an. pigs, 92/10: Pig fat from slaughterhouse waste 17 DK 172775 B1

Feed additive according to Note: The above feed additive mixture

Flex Norm-Mix 2201: Pre-mix of Carbadox (growth promoter), minerals, vitamins and essential amino acids produced and sold by FLEX-FOOD, Randersvej 14, Fausing, DK-8961 Allingåbro.

5

The ingredients, all of which (except the pork fat, which was heated to 50-60 ° C were added liquid) in powder form, were dry mixed by thorough mixing in a snail mixer for 20 minutes. The pre-mixed feed was available as a free-flowing, essentially dust-free powder or microgranule ready for use in an automated 10 feeding plant.

Testing and test results

For a test experiment with weaned piglets, a basic dry feed 15 of the following composition was prepared:

Ingredients dry matter weight% amount% kg 20

Wheat (ground) 87.0 72.00 720.00

Soybean (Cheese (Good) 44%) 87.0 10.00 100.00

Fishmeal, ash low 94.0 10.00 100.00

Fat, an. pigs, 92/10 99.5 4.00 40.00 25 Flex Premix 2292 96.3 4.00 40.00

Total 100.00 1000.00 30 and a dry feed according to the invention having the following composition: 18 DK 172775 B1

Ingredients dry matter weight% amount% kg 5

Wheat (ground) 87.0 69.50 695.00

Soybean (Cheese (Good) 44%) 87.0 10.00 100.00

Fishmeal, ash low 94.0 10.00 100.00

Fat, an. pigs, 92/10 99.5 4.00 40.00 10 Feed additive according to. opf. 90.5 2.50 25.00

Flex Premix 2292 96.3 4.00 40.00

Total 100.00 1000.00 15

Flex Premix 2292 is a premix of Carbadox (growth promoter), minerals, vitamins and essential amino acids produced and sold by FLEX-FOOD (supra).

The dry feed containing 2.50% by weight of the feed additive of the invention contained 0.50% by weight of pure 20 Mimosacea extract.

Both feed mixtures were prepared by dry mixing the ingredients, all of which (except the pork fat, which was heated to 50-60 ° C was added liquid) in powder form, by thorough mixing in a snail-forced mixer for 20 minutes. The fermented blend feed compositions were available as free flowing, essentially dust-free powders or microgranules ready for use in an automated feeding facility.

A total of 69 piglets with an average weight of 6.3 kg were removed on the same day from their mothers origin and were distributed in 6 paths with 35 pigs in 3 control teams and 34 pigs in 3 30 experimental teams.

All 69 pigs were fed the "Cranstart 150" for the first 13 days after weaning (starter feed containing "Olaquindox" as growth promoter and used for 10-14 days for weaners weighing 5.5-6.0 kg and which is supplied by Aller Mølle, Christiansfeld, 19 DK 172775 B1 DK), during the last two days of this period a slow transition to the above-mentioned basic dry food without content of the feed additive according to the invention occurred.

Thereafter, the pigs in the test teams were fed for 15 days with the dry feed mixture containing the feed additive of the invention, while the pigs in the control teams continued with the base dry feed without containing this additive.

The experiment was terminated by feeding both teams for 5 days with the basic dry feed without the content of the additive according to the invention.

10 Throughout the experimental period from weaning to the end of the observation period, all pigs had free access to clean drinking water in drinking cups around the clock, and during the same period (except for the last 5 days during which both control teams and experimental teams were fed the basic feed) drinking water was mixed. 4 96o lactic acid. - If a pig developed diarrhea, it was treated with 1 ml of Potrox® per day. day for two consecutive days, whether it belonged to the control team or the trial team. Treated pigs remained in the trial.

(Potrox® is a sulfo-trimethoprim combination preparation). - All the pigs were weighed before the start of the test and again after the end of the test.

20 Experimental Results

The obtained test results are shown in the following table, which shows that one pig died from the test team during the observation period. It died during the final 5-day period during which both the control team and the test team pigs received basic dry fodder without the feed additive content of the invention. The cause of death is believed to have been 25 toxin shocks due to diarrhea.

In addition, it is seen: 1) that the test team pigs required far fewer diarrhea treatments than the control pigs -12 versus 42 (Most of the diarrhea cases in the test team occurred in pigs in the two paths that were closest to the control teams' paths), 20 DK 172775 B1 2) that the pigs of the test team had on average a better appetite than the pigs of the control team and averaged 0.77 Fes (feed units) per day. day per pigs versus 0.59 Fes. day per 3) that the test team's pigs had on average a better daily growth than the control team's pigs - 379 grams per day. pig per per day versus 267 grams per day. pig per and 4) that the pigs of the test team had on average better feed utilization than the pigs of the control team - 2.08 Fes. kg added vs. 2.28 Fes. kg added.

10

All in all, the experiment shows that by feeding a feed containing an additive according to the invention, very significant benefits are obtained, both in preventing diarrhea in weaned piglets and in promoting appetite, growth and feed utilization. Although diarrhea cases occurred among the pigs fed 15 feeds containing the additive of the invention, the number of diarrhea cases was far fewer than in the control pigs and it is believed that the number of diarrhea cases could be further reduced or completely eliminated by partly avoiding that pigs fed feed containing the additive of the invention walk in paths adjacent to paths of pigs fed non-additive feed and therefore develop infectious 20 diarrhea, and partly by increasing the content of additive according to the invention, so that the feed contains a total of e.g. ca. 1 wt% pure Mimosaceae extract powder.

Finally, it should be noted that the reason why the experiment was started with pigs 13 days after their weaning is that, in the pig farm in question, the weaning-25 crisis with dangerous diarrhea is experienced, approx. 13 days after the ffabning day. In other holdings, the weaning crisis may occur earlier, e.g. 4-7 days after weaning, but also occasionally or even later.

DK 172775 B1

2I

Experimental results obtained by feeding piglets with basic dry feed without (control) or containing feed additive respectively. opf. (trial) average weight per pig per harvest: 6.3 kg average amount of feed per pig per day Before an increased 0.1 »kg average amount of energy per pig per day Before experimental species: 0.23 FEs control control control control Thursday trial trial forteo hold m ___________ 1 2 3 all 4 $ β or

Fravannine__7-i and I | 'P ~~ T 1 f

Number of grays per team at foreeaaatart ___H_t2 (2__35 'Tf ~ fg "Tf J« gannaminltltg guard per qrts at? Or »eqiatart o.o e.f 7.1 a.o fc] o eTf gj 5 ^ FEaprdagprprie --r ~ --—

Test Street; ___ 124ϊ2__0.1 »" "" θ, 1β ____ _____ * 14 * 2 ___ 0ΛΪ 0.38 _______ 22-ian _____ 0.56 "0.77 __ Sj'Jan ____ 0.56 o, 58 -: ---—- 154 * 2 ---- --- 0 * 36 0.66 —-------. 25-Jen ______ 0.33 ------ Qjj -m «--- lir --- sis # ___ 27-Jan _____ 0.36 077 -_§ * i * 2 ------ 0 ^: - —-— tn - ------ & -------- 077 _______ 3yJ * f> __._____ 0.56 '' '~ λ 77 _114 * 2 -___ 0.56 ~ - Oj7 _________Wjtob__ 0 * 36 ~~ —---- Ktv ~. 02-fat> ____ ~~ ό! 64 ------ r? 6 ______ Mbife__5 & ----- si?

Attempt End _____ 04-Feb__0.0e ----— —-- ifef ___- '. -. ghteo____ "- --- ΐ" ο5 ------- & _ ... Mg * ______ j! g · ---- get • - -. - ° 9 ~ ---- 3R ___ 0.60 average number of Fas per grla per day “" ---------- In oertoda with Were * HtdoUna (20/1 · 4/2) α & a ...

_ ____________ '—— J-. -----_--, —Sil? gennamanKHg anUki? ee per pig per day ~ “———---, ------- in the hall« periodtn (20 / t · e / g) ______. ______ 0 ^

Number of pigs wdjorsjgesM _ if 12 12 "as --- rv -rr-κ average guard per pig road seal * 14, s nj ~ Ίϊβ ~ g ^^ glejiiytøk ^ g) UP? Zjg Zjll H | g ^ —TST - 379

Fee per ko tiivakat ^ ~ '7 «" ----— - -------——> £ * = 2, _____ 2.06

Number of dlemab dealers ~ ------------- gvrloe behlfllnaer J · T1 ^ -__- _ 22 DK 172775 B1

Scheme 1

Typical monomeric Mimosaceae extract flavanoids are shown below using standard stereochemical imaging. Arrowheads indicate groups that are above the plane of the paper, and dashed lines indicate groups that are below the plane of the paper.

Flavan-3, 4 diols

Oh 'ifV' (fbr-½ IS 4> '. 4h 0h (+ l-Leucofisetinidin W-Leucorobinetinidin'

Flavanolglycosider

OH

T Π HO O

ftO O ·

Quercetrin Myricitrin

Flavanones o (i) -Butin

Chalconer o «jQf o- jdr o '°

Butein flobtein

Scheme 2 23 DK 172775 B1

Flavan-3-oler "OCX0" -txjfc (-) - Fisetinidol (-) - Robinetinidol

'9Cx0 ·' "ooA

Oh Oh * (♦ l-Catechin (♦ l-Gatlocatechin

Dihydroflavanoler

Ow W01 '~ οχχα:' o (+) - Fu $ tin {♦ J-Dihydrorobinetin

Flavonols fV "A, -

"OCX0" -ffA

0 Oh

ISLAND

Fl ** l! N Robineiin

Scheme 3 24 DK 172775 B1

Main fragmentation products 5 cleavage '

/, '_ o «_.oM

- * HOOC—— ° h iomi Protocatechinic Acid

(OHt / IOM) \ _ / OH

~ -Q- ~ -0- w

ABOUT

Resorcinol Phloroglucinol 15

Scheme 4 20

Smaller fragmentation products cleavage

A .Oh _ / ®H

H ^ C ^ ° "V, ΊΟΗ) N— Catechol

(OHI iomi \ __ / ° H

"N. blast of / (7)) - oh S molecule Ί0Γ ΊφΓ ^

ABOUT

Resorcinol Phloroglucinol DK 172775 B1

Scheme 5

Biflavanoids and triflavanoids from fresh bark samples from Mimosaceae have been shown to have the following structure.

Biflavonoider

Oh-. , ji, C

.mi.

OH OH

A. Pure trans-stereophorins r,, r2 H: (-) - fiselinidol - (+) - catechin R, -. OH, R OH: (-) -robinetinidol - (+) -gallocatechin B. 2,3-trans-3,4-cis: 2 ', 3'-trans stereoforms

Rr Ra H: (-) - physisinidol * (+) - catechin

Scheme 6 20

Triflavonoids c · Γ | - ^ - 5> γΌΗ om i j; h · i HOv ^ S. N ^ A'0h B r Jøuj.

\. ·· N / 'oh i- k

QH

Pure trans stereoforms R OH: | 4.8 ": 4 \ 6") · () -robinetinidol- () -robinetinidol- (i) -gallocatechin RH - (-) - roninetinidol (!) catechin

Five isomeric triflavonoids were isolated

Claims (9)

1. Animal feed additive comprising a flavanoid-containing extract derived from a plant belonging to the family Mimosaceae. characterized in that it further comprises an odor and / or taste masking substance or a mixture of such substances which mask the odor and / or taste of the flavanoid extract, and optionally one or more carriers for the odor and / or taste masking substance or mixture thereof. . Additive according to claim 1, characterized in that it is in powder or micro-granule form or a mixture of these forms. Additive according to claim 1 or 2, characterized in that the flavanoid-containing extract is obtained from the plant Herba Acacia Catechu Willd. preferably the variety Heleatii. Am. or Acacia meamsii De Willd (Black Wattle). 15 Additive according to any one of claims 1-3, characterized in that the content of polyhydroxy-flavans in the crustacean flavanoid-containing Mimosaceae extract constitutes 65-75 wt.% Of the total extract mass, preferably 67-72 wt. . 70% by weight. 20 An additive according to any one of claims 1 to 4, characterized in that it contains from 1 to 99% by weight of flavanoid containing Mimosaceae extract. preferably 5 - 80% by weight, more preferably 10-50% by weight, most preferably 15-30% by weight, in particular approx. 20% by weight, based on the total weight of the additive, while the remainder is an odor and / or flavoring substance, carrier therefor, and any filler and other additives. Additive according to any one of claims I-5, characterized in that the odor and / or taste masking substance is selected from vanilla, vanilla sugar, whey powder, apple cider powder and apple vinegar powder or mixtures thereof. 30 DK 172775 B1 Additive according to any of claims 1-5, characterized in that the odor and / or taste masking substance is provided in the additive in the form of vanilla-containing biscuit flour. Animal feed or beverage, characterized in that it comprises one or more feed intended for the animal concerned and a beverage and an additive incorporated therein according to any one of claims 1 to 7, preferably in such weight ratios that it The pre-mixed feed or beverage, respectively, comprises 0.05 -10% by weight of the flavanoid-containing extract, more preferably 0.1-5% by weight, most preferably 0.5-2% by weight, in particular approx. 1% by weight, calculated on the basis of the weight of the total feed or drink mixture. Process for the preparation of a feed or beverage for controlling Salmonella typhimurium DT 104 in an animal, characterized in that a feed or drink intended for said animal is added to an additive according to one of claims 1-7, preferably in the weight mixing ratio specified in claim 8.