WO1999045795A1

WO1999045795A1 - Additive for animal feed containing flavanoids from a mimosa-extract and animal feed containing the same and a process for producing the animal feed

Info

Publication number: WO1999045795A1
Application number: PCT/DK1999/000120
Authority: WO
Inventors: Georg Andresen; Hans Jensen
Original assignee: Diarex V/G. Andresen; Flex Foder Holding Aps
Priority date: 1998-03-12
Filing date: 1999-03-10
Publication date: 1999-09-16
Also published as: DK172775B1; DK33598A; AU2713499A

Abstract

An animal feed additive, which comprises a flavanoid containing extract gained from a plant belonging to the Mimosaceae family and furthermore comprises a smell and/or taste masking compound or mixture of such compound, which mask the smell of the flavanoid extract and/or taste, and possibly one ore more carriers for the smell and/or taste masking compound or the mixture hereof, has appeared to be applicable in practice for fighting against and/or prevention of diarrhoea by animals, because the animals with big pleasure and eager voluntarily will eat a feed or a drink, which contains the additive, which is not the case with the feed, that contains the pure Mimosaceae extract, even in very little concentrations. The animal feed additive can thus be utilized instead of the traditional antibiotics, which are used for the above purposes and as growth promoters.

Description

Additive for animal feed containing flavanoids from a mimosa-extract and animal feed containing the same and a process for producing the animal feed

The present invention relates to an additive for animal feed which encompasses an extract with flavanoids that are obtained from a plant belonging to the Mimosaceae family and an animal feed containing such additive. The additive can be given directly to animals which are in a need therefor, but is particularly intended for admixing into a feed or a drink intended for the animal/ s concerned. The additive and feed or drink containing this is able to fight against and/or prevent diarrhoea and other gastrointesti- nal tract disturbance or suffering in animals, increase the growth of animals and/or increase the feed yield in animals which are fed with the additive or the feed or drink containing the additive.

The additive and the additive containing feed is particularly intended for production animals within the agricultural sector, such as pigs, cattle, sheep, goats, horses and fowls and other poultry and their progeny, such as sucking pigs, piglets, calves, lambs, kids, foals and chickens etc. but is also usable to pets, such as dogs and cats and their progeny.

Background of the invention

In the latest decades in the agriculture production of domestic animals a steadily increasing rationalisation, specialisation and intensivation has taken place so that on the individual agricultural enterprise today normally only a few persons are employed, yes, often only a single person, so that the demand of a high productivity and short production time is extremely big, so that in most cases a production of only a single animal species is taken place, e.g. porkers, broilers or meat cattle or products from this animal species, e.g. piglets from sows, milk from cows or eggs from egg laying fowls. In return of this, the animal species in question is kept in a very large number from several houndreds to several thousands individuals on the individual agricultural en- treprise. Firstly, these measures cause that almost all care and feeding of the animals must be automatized and may be carried out mechanically, secondly, that the animal constantly are exposed to a big pressure of infection due to the many animals of the same species on one enterprise and thirdly that the physiological systems of the animals, i.a. the digestive system, due to the demand of high productivity and short production time constantly will be exposed to a big pressure so that even minor extra loads can make the system break down which for the digestive system, e.g., can result in violent diarrhoeas which either cause that the animal do not thrive or possibly after shorter or longer time purely and simply die, which in both cases implicates loss of production for the farmer.

The demand of a high productivity and a shorter production time are also expressed by the fact that the progeny of the animals, e.g. piglets and sucking calves are weaned, i.e. are removed from their mother animals and thereby from their former natural feed, the mother milk, at an earlier time in their lives than previously was the case, which for the young animals was a physiological more appropriate point of time in their growth and maturing toward the independent level for a young animal. Thus, today piglets are weaned normally at an age of only 4 - 5 weeks after the birth contrary to 8 - 9 weeks previously.

The early weaning of e.g. piglets and the transition to exclusively other feed than the mother milk of the sow leads inevitable to an immediate or after short time entering into a cronical condition of diarrhoea by almost all piglets and without treatment these pigs do not only thrive for a long time thereafter but in many cases simply die.

The above relationship has been known as an envitably consequence of the production development in the agricultural enterprises, and when this nevertheless has been able to take place it is only because of a concurrent still more widespread use of antibiotics of various kinds to the animals either given as parentel injections into the animals or what is more common admixed to the feeding of the animals. In the latter case the additions of antibiotics in question are usually mentioned "growth promoters". Antibiotics applied to the animals, e.g. growth promoters, have, however, a tendency to provoke resistance of the bacteria present in the animals and other microorganisms, and it has therefore been necessary constantly to develop and use new antibiotics to the animals. Some of the used antibiotics to the animals are, however, the same or chemically so closely related to those antibiotics which are used in the fight against deseases provoked by bacteria and other microbes in humans that in the public opinion, especially in the most recent years there has been an increasing concern that the resistance which is found in the microorganisms in the animals should be transferred to the desease provoking microorganisms living in human beings, whereafter these will not be fightable with the present human antibiotics at disposal.

On this background there is therefore a continuously increasing demand to limit the use of antibiotics of any kind to animals in the most possible way and completely avoid, yes, directly forbid the use of antibiotics of any kind as "growth promoters" in the animals feeding.

From the above explanation of the production conditions in todays agricultural enterprises it will be understood that such a prohibition against growth promoters obviously will cause catastrophic conditions for both the animals, the production results and the economy of the agricultural enterprises, unless alternative treating methods or measures are provided in advance which are not based upon compounds of antibiotic character and therefore for the time being large search and research are taken place after such method and measures, both after quite new as well as after known and previously used ones, which for some reason are taken out of use either during or before the "period of the antibiotical growth promotors".

Prior art

In WO patent publication No. 96/03150 compositions of hemi cellulose fibers and polyphenols are described which are extracted from larch threes (Larix occidentalix Nuttaiπ. The exemplified hemi cellulose fibers and polyphenols consist thus of the water soluble arabinogalactan respectively taxipholens. These compositions are stated to have a growth promoting effect on beneficial bacteria, such as Bifidobacterium thermophilum and an inhibiting effect on maleficial, patogenic putrefactive bacteria, such as Clostridium ssp.. Bacteroidaceae and Candida ssp.. in the gastrointestinal tract with human beings and animals and thereby fight against gastrointestinal disturbances, such as diarrhoea, with these. It is mentioned that flavanoids and tannins are within the group of polyphenols which should be usable in the mentioned compositions but this is not either exemplified or in any other way verified. The relation between fibres and polyphenols in the compositions are given to be between 20 and 3 and such compositions are mentioned to be added to the animal feeding in an amount of 0,1 - 5 percent by weight, preferably 0,1 - 2 percent by weight, based upon the weight of the animal feed. However, in the publication tests are described only with the clean arabinogalac- tan or clean compositions of arabinogalactan and polyphenols extracted from larch trees in connection with testing of the effect on various Bifido-bacteria species grown in a test tube containing a fortified clostridial agar broth but no forms of experiments of feeding of animals with the feed which contains arabinogalactan-polyphenol com- positions are mentioned. It is thus not known if the composition or a feed which contains this will be effective in practice or if there e.g. will be problems getting the animals voluntary to eat such feed. Moreover, it is mentioned that the compositions can be formulated to tablets, pills, capsules, pastils and syrup and that these dispensing forms may contain sweetenings, such as sucrose or saccharin and flavour additives, such as peppermint, methylsalicylat or orange aroma. These dispensing forms are normal for human beings but not for animals and nothing is described that these should be admixed or in any other way intended for admixing to an animal feeding.

Another known drug, which before the discovery of the antibiotics and especially be- fore the advent of the growth promoters have been used in the agricultural enterprises for fighting against and/or prevention of diarrhoea conditions in animals especially in sucking pigs and piglets, sucking calves and other young animals 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 Aid" and later on under the name of Diarex^® in a form of either a mixture which beyond water and a small amount of preservative contained about 50 percent by weight of the flavanoid containing plant extract or in form of a powder which apart from a minor amount anticlotting remedy consisted of the clean flavanoid containing plant extract. The drugs were given to the animals directly as they were as purchased in the recommended dose (2 - 30 ml or g per animal per day) or they were admixed the animal feed or drinks.

However, the Mimosaceae-extract has for the animals a repulsive smell and/or taste and the animals will therefore normally under no conditions eat the extract voluntarily, no matter if it exists as a mixture, powder or is admixed to their feed or drinks. Earlier it was necessary that each individual animal during the feeding was held by one or more persons while a second (or third) person with a syringe or feed- or powder gun injected the mixture/the drink respectively squeezed the powder or the feed to which the mixture or powder were added directly into the gullet of the animal. By small animals, such as sucking pigs or sucking calves, it was a feasable although a very laborious and time consuming task to threat each individual animal up to more times daily in this way, but by larger animals it was almost impossible due to the physical size of the animal and its strength, unless special accomodations were applied for fixing and locking the animal in an unmovable position.

The treatment of the prevention of diarrhoea conditions of the animals by means of the Mimosaceae-extract in the above described way was by itself effected and inexpensive but due to the labourious and time and working power demanding way of feeding it ran nevertheless practically completely out of use because antibiotics and growth promoters entered the market at economically feasible prices because these compounds did not have for the animals any repulsive smell and/or taste and therefore could be admixed directly into the usual feed of the animals which feed the animals would like to eat voluntarily.

In the present situation where the use of the antibiotics as growth promoters and diarrhoea fighting and preventing remedies to production animals expectedly must be given up completely within a near future in a continuously increasing number of countries the inventors up to the present invention have discovered that it would be desirable if possible to change the character of the Mimosaceae-extract so much and or develop an administration method for this which is not connected with the above mentioned difficulties as a consequense of the animals not voluntarily drinking or eating the extract containing mixture/drinks respectively powder/feed, because the Mimosaceae-extract does not contain compounds of an antibiotical character and as a whole is not absorbed from the gastrointestinal tract in the organism of the animals.

The first time it was thus tried to admix the Mimosaceae-extract to the normal feed of the animals in such a little concentration that the animals were not repelled by smell and/or taste and therefore would like to eat the feed voluntarily, but as a consequence of the animals' very fine and well-developed smelling and tasting sense it appeared that the low concentration required herefore was so little that a sufficient therapeutical or prophylactic effect was not achievable by the amount of feed voluntarily eaten by the animals.

Thereafter, it was tried to encapsulate the Mimosaceae-extract powder as coated par- tides or granules which then were admixed to the feed of the animals but again because of the animals' fine sense of smell and/or sense of taste neither this method gave a satisfactory result for the same reason as mentioned above. Besides, the coating process implies that the product became too expensive for application as a normally used feed additive in practical breading and production of animals.

Brief description of the invention

Therefore, it is very surprising that it with the present invention nevertheless has been made possible to provide the Mimosaceae-extract in a special and up till now unknown additive form which makes it possible to get the animals such as for instance weaned pigs and calves to voluntarily eat the additive form either directly as it is or admixed to the animals usual feed or drinks in so sufficient amounts that a therapeutic and prophylactic effect can be obtained especially regarding the fighting against and prevention of diarrhoea conditions of the animals.

According to the invention this is obtained with the feed additive which comprises a flavanoid containing extract gained from a plant belonging to the Mimosaceae family, which feed additive is special by the fact that it furthermore comprises a small and/or a taste masking compound or a mixture of such compounds which mask the flavanoid containing extracts smell and/or taste and possible one or more carriers for the smell and/or taste masking compound or the mixture hereof.

Apart from the above components the additive can also contain other ingredients, such as real filling up compounds and/or means for extension, dyes, consistence conditioning compounds, essentiel minerals, vitamins, amino acids etc., which do not repel the animals or decrease their appetite or eager to eat.

Preferred embodiments for the additive according to the invention are stated in the dependend claims 1-7.

The following remarks are made for the invention and the embodiments for said invention: The feed additive according to the invention will mostly be present in a form of dry powder or minigranules but will possibly also be present in humidified, pasta-like or liquid form. Irrespective of the physical condition the additive can generally be prepared by simple, thorough mixing of the components. Thus, a powder additive can be prepared by dry-blending of the ingredients, but it can also be prepared by wet- blending of the ingredients followed by drying, grinding and possible sifting. A mini- granulate can be prepared the same way although to begin with possibly sufficient amount of liquid is to be added of an appropriate kind (e.g. water) in order to obtain sufficient adhesion power between the particles of the ingredients.

By a smell and taste masking compound is to be understood any not health damaging compound which in a sufficient concentration is able to mask for the animals the unpleasant and repulsive smell and/or taste from the Mimosaceae-extract to such a degree that the animals voluntarily will eat sufficiently enough of the additive or a feed/a drink in which the addive is admixed so that the animals are eating a therapheutical and/or prophylactical amount of the Mimosaceae-extract. The smell and/or taste masking compound can exert its masking effect in different ways. For instance it can: 1) by itself have such a pleasant and attractive smell and/or taste which drowns or is mixed with the smell and/or taste from the Mimosaceae- extract so that the latter either is not conceived or is not repulsive against the animal, 2) exert a blocking effect on the receptors in smell and/or taste cells of the nose and mouth of the animals which nose and mouth register the Mimosaceae-extract as smelling and/or tasting unpleasantly,

3) bind itself physically/chemically to the compounds or molecule groups in compounds of the Mimosaceae-extract which cause the repulsive smell and/or taste on the animals and thereby block their smell and taste effect, assumed through that the Mimosaceae-extract is not hereby loosing it therapheutical and prophylactic effect, or

4) have a random combination of the above properties or effects.

Examples of the smell and/or taste masking compounds which exert their masking effect upon one or more of the above ways are mentioned in claims 6 and 7. Especially preferred, in this connection, is an additive which contains 5 - 80 % by weight of a flavanoid containing Mimosaceae-extract and 95 - 20% by weight of biscuit meal, preferably 10 - 50% by weight of extract and 90 - 50 by weight of biscuit meal, more preferably 15 - 30 % by weight of extract and 85 - 70%) by weight of biscuit meal, particularly about 20%) by weight of extract and about 80%) by weight of biscuit meal based upon the total weight of the additive.

The possible incorporation of one or more carriers for the smell and/or taste masking compound in the feed additive according to the invention depends upon which specific compound is in use and the aim of its incorporation will accordingly be dependend upon this. To certain of the very strongly smelling and/or tasting masking compounds it will be necessary with a carrier for simply to achieve the sufficient solution of the former in order to obtain an appropriate dosing and homogeneous admixing with M mosaceae-extract. For other masking compounds it will be necessary with a carrier in order to maintain the masking compound intact within the additive mixture, e.g. to antagonise the evaporation, separation, break down or other harmful alteration of the compounds during the storage of the additive for a shorter or especially a longer period of time. In other cases the function of the carrier can be to provide a so sufficiently tight binding of the masking compound with the evil-smelling or evil-tasting compounds of the Mimosaceae-extract that the desired masking effect will be achieved.

As carrier can be applied any not unhealthy compound which is able to establish the above effects and which has no harming influence upon the therapheutical and prophylactic effect of the Mimosaceae-extract in addition to that it of course not by itself at the same time may have any smell and/or taste which will have a repulsive effect upon the animals. The carrier can be of organic or inorganic origin, digestible or indigestible. As examples of carriers can be mentioned carbohydrates, such as starch particles, cellulose fibres and mono-, di- and oligosaccharides, proteins, e.g. soya protein, fat and oils, and salts, such as calcium phosphate and sodium chloride, and as well as mixtures of such compounds. As an example of compounds which usually not can be used as carrier can be mentioned iron containing compositions of compounds of various kinds as the iron in these compounds in many cases will block the effect of the therapheutic and prophylactic effects of the Mimosaceae-extract.

That the additive is present in powder form means that the biggest diameter of the substance particle of the additive does not exceed about 0,2 mm. That the additive is present in a form a micro granulate means that the main part, e.g. at least 80% by weight, mostly at least 90% by weight, more preferably at least 95%> by weight and most preferably at least 98% by weight, of the particles have a largest diameter, which is in the area of 0,2 - 2,0 mm.

Relating to the flavanoid containing extract per se this is gained as mentioned above from a plant belonging to the Mimosaceae family. These plants are tree or bush-like plants which inter alia grow in the southern part of Africa where they are grown in various places in the plantation which are harvested when the bark of the plant is considered to be mature for gain of the flavanoid containing compounds herein. For the tree-like plants this means when the plants usually are at least 9-10 years old. Usually, the harvesting takes place in the period of growth of the plants that is to say in the months of September to March where the bark has a humidity of about 50%. The bark is taken off the plants and the bark is transported, possibly after a preceding drying to bark bars, to extraction facilities in the country or abroad, where it is cut into shavings which are subjected to an autoclavation extraction process with water at superatmos- pheric pressure and at a temperature over 100°C or an extraction with percolation with water at a lower temperature after rupture of the parenchymal cells of the bark by autoclavation. After concentration of the extracted liquid spray-drying is taking place in a countercurrent spray dryer for procurement of a redly, yellowly, or greenly-brown powder with a humidity of about 6%.

The powder, which consists of hollow, spherical particles, has a density of 370 til 500 kg/m , preferably 370 to 430 kg/m , is slightly hygroscopic and is soluble in for instance water and alcohol. A 13% solution of the powder in water has a pH of 4.6, a minor buffer area between pH 2.8 - 5.8 and a bigger buffer interval at pH 9.5. Aqueous and alcoholic solutions are stable by long time storage in the dark, but its expona- tion to sunlight will develop red colouring. The powder is by dry and dark storage at room temperature stable for at least 3 years. By exponation to sunlight a red colouring of the surface layer will take place, and by heating a commencing carbonization will take place at temperatures above about 230°C. The content of polyhydroxyflavanoids in the powder depends of the specific Mimosaceae species, from which the extracted powder is gained, but will usually be of 65-75 % by weight, particularly 67-72 % by weight, and particularly about 70% by weight. By mixing of the two productions an egalization can take place in the area of 67-72 %.

An alcoholic (99.8 ethanol) extraction of powder will in a 0.25 % (weight/volume) solution by addition of 0.25 ml of 1% FeCl₃ to a 10 ml extraction solution result in a clear blue solution without precipitation. The presence of tannic acid will give a blue precipitate after centrifugation and should not occur.

Thin layer chromatography on silica plates of the alcoholic (99.8 % ethanol) extraction of the powder developed with the butanol/acetic acid/water (dest.) in the mixing relationship 4: 1 :5 renders R_f values, which depend upon the special Mimosaceae species, from which the extraction powder is gained, and possibly also upon the extraction conditions. Extraction powder from Acacia mearnsii de Willd. which grows as trees in South Africa, and which is gained by autoclavation extraction with water at about 1 10°C for about 6 hours, has thus an R_f of 0.70, while extraction powder from

Herba Acacia Catechu Willd. species Helgati. Arn.. which grows as bushes or plants in Southwest Africa (Namibia), and which is gained by water percolation for 48 hours after autoclavation rupture of the parenchymal cells, have an R_f of about 0.35. These values must be considered in relation to that tannic acid under the same conditions has an R_f of 0.9.

Other physical and chemical characteristics of the Mimosaceae extraction powders are to be found in the special literature.

As mentioned 65-75 % by weight of the Mimosaceae extraction powder is made up by polyhydroxiflavanoids, while the rest is made up by a complex mixture of carbohydrates, gummies, sugars, amino acids, and other small components, from which a little amount is insoluble compounds. The part, which is made up by polyhydroxy flavanoids, consists of condensed flavanoids and their monomeric precursors. Polyhydroxy flavanoids constitute thus a complex mixture of a number of various single components, which basic unit is the typical carbon- 15 flavanoid skeleton, and their molecular weights vary from about 300 for the monomeric precursors to about 3000 for the biggest polymers with an average molecular weight of about 1200-1300. The average Mimosaceae flavanoid molecule consists consequently of 4-5 monomeric flavanoid molecules. They are bound to each other with carbon-carbon bonds. The flavanoid precursors consist typically of flavan-3.4-dioles (leucoanthocyanidins) and flavan-3- oles (catechins), reference is made to overleaf tables 1 and 2.

By disintegration of the flavanoid molecules the heterocyclic C-ring is typically rup- tured followed by production of resorcinol, phloroglucinol and protocatechinic acid, gallic acid and in smaller amounts resorcinol, phloroglucinol and catechol, pyrogallol, reference is made to overleaf tables 3 and 4. The polymeric flavanoid molecules consist prevailingly of flavan-3.4-dioles condensed with flavan-3-oles. The structure of the di- and triflavanoids are shown overleaf in tables 5 and 6.

The polyflavanoids have a strong affinity to collagen, which forms the basis for their application in the traditional, vegetarian tanning methods. The true affinity of the flavanoid substances to proteinaceous material was first clear by the time for the level of triflavanoid, which can be seen by the finding that gelatine is precipitated from a triflavanoid solution under controlled conditions. Further information about the chemical composition of the Mimosaceae flavanoids and the stereostructure and about the contents of non-flavanoid substances in the Mimosaceae extracts are to be found in the special literature.

The Mimosaceae extracts per se have no direct bactericid or microbiocid effect, that is to say they have no direct killing effect upon bacteria and other micro-organisms. The effect of the extracts on the micro-organism flora (bacteria and other micro- orgamisms) in the gastrointestinal tract by animals and human beings is considered to appear by a precipitation of and binding together of proteins from the outer surface of the intestinal epithelium and in the content of the intestine and in the surfaces of the micro-organisms. Binding and packing of the bacteria and other micro-organisms and especially their toxins, virtually in a protein flavanoid cartridge or network, is taken place and their harmful effect upon the intestinal wall is hindered or decreased essentially, and their number within the intestine is decreased very much by the peristaltic exhausting from the intestine of the indigestible protein flavanoid cartridge or complex. As the micro-organisms are not directly killed their toxins are not released to a big extent it happens by application of big doses of traditional antibiotics, which can lead to a toxin shock and the death of the treated individual. Furthermore, non resistent bacteria species are produced by the flavanoid treatment, because the fighting is based upon a general, physical-chemical catch and binding of the micro-organisms which then are excreted with faeces instead of at a specific chemical/biochemical intervention into the life and/or reproduction processes of the organisms against which surviv- ing organisms will be able to develop measures toward and consequently future generations will become more resistent to the operational mode of the pertinent antibiotics.

Following administration of a single dose of the Mimosaceae extract in some cases such a big number of bacteria/microorganisms may still be left in the gastrointestinal tract that they will rapidly propagate and thereby continue or start up again a condition of diarrhoea in the treated individual. The Mimosaceae extract should therefore normally be administrated at least two times dayly for 2-4 days and possibly longer in order to achieve optimal results.

The effective components in the Mimosaceae extract are not directly digestible, and neither these nor the breakdown products hereof are detectable in the blood or in the milk by individuals, who orally have been given the extract. Oral administration of the extract into an individual results therefore only in a local effect in the gastrointestinal tract for what reason systemic adverse events hardly exist and therefore they have not been reported. Furthermore, there will be no need for "retention times" for meat or milk from animals, which have been treated with the Mimosaceae extract.

Because the Mimosaceae extract alone is effective in the gastrointestinal tract by purely physically to decimate the amount of microorganisms [such as: A) anaerobic gramnegative rods (bacteroids), cocci (pepto-streptococci and peptococci), clostridies and lactic acid bacteria; B) aerobic (about 1 % of total) enterobacteria, streptococci species and bacillus species; and C) various fungi etc.] antagony the tendency to over- growth of one or a few species which are often seen following use of antibiotics.

Apart from an animal feed additive, which comprises the above mentioned described flavanoid containing Mimosaceae extract, the invention also relates to an animal feed or drink, which is particular by comprising a feed or drink, respectively, aimed at the pertinent animal and a mixed additive according to the invention, preferably in such weight relations that the ready mixed feed or drink, respectively, comprises 0.005 - 10 % by weight of the flavanoid containing extract, more preferably 0.1 - 5 % by weight, most preferably 0.5 - 2 % by weight, especially about 1% by weight, calculated on the basis of the weight of the feed or drink mixture in total.

The additive according to the invention can be used for treatment and/or prevention of diarrhoea or other gastrointestinal tract disturbance or suffering in .an animal, promotion of growth of an animal and/or increase of the feed yield in an animal, which method is particular by giving the animal a feed or a drink according to the invention.

The additive according to the invention can be used for fighting against Salmonella typhimurium DT 104 in the gastrointestinal tract of animals, by giving the animal an additive, a feed or a drink according to the invention.

Finally, the invention relates to a method for preparation of a feed or a drink for fighting against Salmonella typhimurium DT 104 in the gastrointestinal tract of ani- mals, which method is particular by admixing an additive to a feed or drink intended for the pertinent animal according to one of the claims 1-8, preferably in those in claim 9 given mixture relationships of weight.

As by the treatment with the pure Mimosaceae extract it holds true by treatment with the animal feed additive or with the additive containing feed or drink that following administration of a single dosis of the feed additive or the additive containing feed according to the invention there will possibly still could be such a big number of bacteria or microorganisms left in the gastrointestinal tract in a number of cases that they will quickly be propagated and thereby continue or restart the condition of diarrhoea in the treated individual. The feed additive or the additive containing feed according to the invention therefore should normally be administrated at least twice dayly for 2-4 days and possibly longer to obtain optimal results. The big advantage by the application of the feed additive and/or the feed/drink according to the invention is, however, that while the pure Mimosaceae extract must be administrated to the animals by com- pulsory feeding then the animals voluntarily and with big pleasure and eager consume the additive and/or feed drink according to the invention, and thereby achieve the ad- vantageous, therapeutic and prophylactic effect without application of the time consuming and expensive working power efforts.

By feeding animals with the additive, the feed or drink according to the invention it will thus be possible to prevent or fight against any condition of diarrhoea or other gastrotestinal disturbance by animals and fight against or prevent the presence of any undesired bacteria or other microorganism in the gastrointestinal tract of the animals, including for instance bacteria belonging to the families E.Coli. Salmonella (e.g. S.choleraesuis and S .typhimurium). Clostridium (e.g. Cl.perfrangens . Camphylobac- ter, Serpulina (e.g. S.hvodysenteria and S.pilosicoli and Lawsonia (e.g.

L.intracellularis .

The invention will furthermore by illustrated more closely in form of the following example and test results.

Example

Animal feed additive was prepared according to the invention with the following composition:

Ingredient amount weight% kg

Biscuit meal 400.00 80.00

Diarex^®-powder, cone. 100.00 20.00

Total 500.00 100.00

Diarex®-powder, cone, is a Mimosaceae extract powder containing about l%o .anti- clotting remedy. 16

The biscuit meal, which is prepared and sold by the Internationale Handelsmaat- schappij Robi bv, Holland, is especially intended for young animals and consists of a mixture of 70% by weight of minced biscuit and confectionery side products and 30% by weight of bread meal. The product, which is easy-flowing, is easily digestible, well-tasting and has a pleasant vanilla sugar-like fragrance.

Biscuit meal is stated to have the following composition and feed properties:

Ingredient Analysed Guaranteed content, % content. , %

Humidity 10.6 max 11

Raw protein 9.3 min 9

Raw fibres 0.8 max 1.5

Ash 2.2 max 3

Sugar + glucose min 18

Starch (preboiled) 38

Starch, total 40

Raw fat min 7

Carbohydrates 70.0 67.5

Calcium 0.5

Chlorine (as NaCl)

1.2

Feed value

Feed units per 100 k g: 132.8

EFOS pigs: 96.9

Amino acids

Lysine: 2.2 g/kg

Methionin + cystine 3.7 g/kg Granulometry 98 % max 2 mm

Heavy metals: absent

Microbiological quality

Entero-bacteria per gramme < 1000

Salmonella per 25 gramme absent

Biscuit meal and Diarex® powder both were present in powder form, were drymixed by thorough mixing in a Nauta blender (conical compulsive blender) for 15 minutes and were then available as a dustless (weakly humid/coherent, but otherwise essentially easy-flowing) powder/minigranulate, which was ready for admixing to an ani- mal feed or an animal drink.

From the prepared feed additive a dry feed was prepared for small weaned pigs (age 4- 5 weeks) with the following composition:

Ingredients dry matter weight% amount

% kg

Wheat (minced) 86.0 51.50 257.50

Biscuit meal 89.4 10.00 50.00

Fish meal, low content in ash 94.0 10,00 50.00

Soyaprotein, cone. HP 300 92.0 5.00 25.00

Flex Combimilk STD. 97.0 10.00 50.00

Fat, an. pig, 92/10 99.5 3.50 17.50

Feed additive iflg.opf. 90.5 5.00 25.00

Flex Norm-Mix 2201 96.7 5.00 25.00

Total 100.00 500.00

In the table indicates:

Flex Combimilk STD: Skimmed milk powder added vegetarian fat Fat, an. pig, 92/10: Lard from slaughterhouse offal Feed additive iflg.opf.: The above stated feed additive mixture Flex Norm-Mix 2201 : Premixture of Carbadox (growth promoter), minerals, vitamins and essential amino acids, which are prepared and sold by

FLEX-FODER, 14 Randersvej, Fausing, DK-8961 Allingaa- bro, Denmark.

The ingredients, which all (apart from the lard, which following heating to 50-60°C was added in liquid form) were present in powder form, were drymixed by thorough mixing in a spiral compulsive blender for 20 minutes. The finally manufactured blended feed was present as an easy-flowing, essentially dustless powder or micro- granulate, which was ready to use in an automated feed plant.

Testing and test results

For a test experiment with small, weaned pigs a basic dry feed was prepared with the following composition:

Ingredients dry matter weight% amount

% kg

Wheat (minced) 87.0 72.00 720.00

Soygrits (oast(Bra)44 %) 87.0 10.00 100.00

Fish meal, low content of ash 94.0 10.00 100.00

Lard, an. pig, 92/10 99.5 4.00 40.00

Flex Premix 2292 96.3 4.00 40.00

Total 100.00 1000.00

and a dry feed according to the invention with the following composition:

Ingredients dry matter weight% amount

% kg

Wheat (minced) 87.0 69.50 695.00

Soygrits (oast(Bra)44 %) 87.0 10.00 100.00

Fish meal, low content of ash 94.0 10.00 100.00

Lard, an. pig, 92/10 99.5 4.00 40.00

Feed additive, iflg.opf. 90.5 2.50 25.00

Flex Premix 2292 96.3 4.00 40.00

Total 100.00 1000.00

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

The dry feed, which contains 2,50 weight% of the feed additive according to the invention, contained 0,50 weight% pure Mimosaceae extract.

Both feed mixtures were prepared by dry blending of the ingredients, which all (apart from the lard, which following heating to 50-60°C was added in liquid form) were available in powder form by thorough mixing in a spiral compulsive blender for 20 minutes. The finished manufactured feed compositions were available as easy- flowing, essentially dustless powders or microgranulates which were ready to use in an automated feed plant.

Altogether 69 sucking pigs with an average weight of 6.3 kg were on the same day removed from their maternal progeny and were distributed to 6 pigsties with 35 pigs in 3 control groups and 34 pigs in three test groups. All 69 pigs were the first 13 days following weaning fed with the "Cranstart 150" (starting feed, which as growth promoter contains "Olaquindox" and is applied for 10- 14 days to small pigs with a weight of 5.5-6.0 kg, and which is delivered by Aller Møller, Christiansfeld, in Denmark), and within the last two days of this period a slow transition to the above stated basic dry feed without content of feed additive according to the invention took place.

After this the pigs in the test groups were fed for 15 days with the dry feed mixture, which contained the feed additive according to the invention, while the pigs in the control groups continued with the basic dry feed without content of this additive.

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

During the whole test period from weaning to the termination of the observation period all pigs had free access to pure drinking water in drinking cups 24 hours a day, and in the same period they were (except for the final 5 days, in which both control groups and the test group were fed with the basic dry feed) offered drinking water admixed 4%o lactic acid. - If a pig developed diarrhoea it was treated with 1 ml Po- trox® per day for two consecutive days, regardless if it belonged to the control group or to the test group. Treated pigs remained in the experiment. (Potrox® is a sulfo- trimethoprim combination drug. - All pigs were weighed before the start of the experiment and again after the termination of the experiment.

Experimental results

The achieved experimental results appear from the tabulation overleaf from which it is seen that from the test group one pig left during the course of the observation period. It died in the terminating part of the 5-days' period, in which the pigs from both the test group and the control group got the basic dry feed without content of the feed additive according to the invention. The cause of death is meant to have been toxic shock as a consequence of diarrhoea. Apart from that it is seen

1 ) that the pigs of the test group demanded much fewer treatments for diarrhoea than the pigs of the control group - 12 versus 42 (most of the cases of diarrhoea by the test group appeared moreover by the pigs in those two pigsties, which were closest to the pigsties of the test groups),

2) that the pigs of the test group averagely had a better appetite than did the pigs of the control group and on average they ate 0.77 FEs (feed units) per day per pig versus 0.59 FEs per day per pig,

3) that the pigs of the test group averagely had a better dayly growth increment than did the pigs of the control group - 379 gramme per pig per day versus 267 gramme per pig per day, and

4) that the pigs of the test group averagely had a better feed yield than did the pigs of the control group - 2.08 FEs per kg growth increment versus 2.28 FEes per kg growth increment.

Altogether the experiment shows thus that by feeding with a feed containing an additive according to the invention very considerable advantages are obtained both with respect to prevention of diarrhoea in weaned small pigs and with respect to promotion of the appetite, growth, and yield of feed. Hence, cases of diarrhoea did appear among the pigs, which we fed with feed containing the additive according to the invention, but the number of cases of diarrhoea was much fewer than by the control pigs, and it is anticipated that the number of cases of diarrhoea could be furthermore reduced or totally eliminated by partly avoiding that the pigs fed with feed containing the additive according to the invention are in pigsties which are close to pigsties with pigs, who are fed with not-additive containing feed, and which therefore develop infectious diarrhoeas and partly by increasing the content of the additive according to the invention, so that the feed altogether contains for instance about 1 weight% pure Mimosaceae extract powder.

In conclusion it should be noted that the reason to commence the experiment with pigs 13 days after their weaning is that in the pertinent pig enterprise the weaning crisis sets in with dangerous diarrhoeas notoriously about 13 days after the weaning day. In other enterprises the weaning crisis may set in earlier, for instance 4-7 days after weaning, but also inbetween or even later.

23

Achieved test results by feeding of piglets with basic dry feed without (control), respectively with content of feed additive acc.inv. (experiment)

average weight per pig by weaning 63 kg average amount of feed per pig per day prior to test start 019 kg average amount of energy per pig per day prior to test start 025 FEs

control control control control test test test test

Group no. 1 2 3 all 4 5 6 all

Weaning 7 Jan

Number of pigs per group at test start 11 12 12 35 11 12 11 34

Average weight per pig at test start 90 81 71 80 100 81 97 92

FEs per day per pig

Test start 20 019 019 Jan

21 Jan 019 039

22 Jan 058 058

23 Jan 058 058

24 Jan 038 058

25 Jan 038 077

26 Jan 038 058

27 Jan 056 077

28 Jan 058 077

29 Jan 058 077

30 Jan 056 077

31 Jan 056 097

OlFeb 038 077

02Feb 094 116

03Feb 094 077

Test terminated 094 097 04Feb

05Feb 094 100

06Feb 094 120

07Feb 094 100

08Feb 075 100

09Feb 058 080 average number of FEs per pig per day in period with Diarex addition 059 077 (20/1-4/2) average number of FEs per pig per day in the whole period (20/1-9/2) 061 079

Number ot pigs at test termination 11 12 12 35 11 11 11 33 average weight per pig at test termi145 137 128 136 188 162 168 172 nation average davly growth increment (g) 262 287 271 267 410 386 338 379

FEs per kg εrowth increment 228 208

Number ot diarrhoea treatments 42 12

Other treatments 2

arthπts 24

Table 1

Typical monomeric Mimosaceae extract flavanoids shown below, where standard stereochemical depiction is utilized. Arrow heads show groups being positioned above the plane of the paper and broken lines show groups below the plane of the paper.

Flavan-3, 4 dioles

OH ^'

OH

HO. o

'3*0 H I

| 5 I H ^yy OH

O H..

(+)-Leucorobinetinidin

(+)-Leucofisetinidin

Flavanolglycosides

OH

HO OH

Rhamnose

Flavanones

HO

Chalcones

HO

25

Table 2

Flavan-3-oles

OH

HO O"

OH

HO

(-)-F isetinidol (-)-Robinetinidol

OH H

OH

OH HO..

OH

OH OH

(+)-Gallocatechin

Dihydroflavanoles

OH

-^QC .

(+)-Dihydrorobinetin

Flavonoles

OH

HO

O

Fijetin

Table 3

Main fragmentation products

cleavage

, OH

HO HOOC — < B >-OH acid

of HOOC OH

OH

OH OH

Gallic acid

O H

Resorcinol Phloroglucinol

Table 4

Minor fragmentation products

cleavage

HO

splitting of - OH molecule

OH

HO^ ,OH ,OH

Pyrogallol

O H

Resorcinol Phloroglucinol 27

Table 5

Biflavanoides and triflavanoides from fresh bark test specimens from Mimosaceae are demonstrated to have the below configuration.

Biflavonoides A.

OH OH

A. Pure trans stereoforms

R,^' R₂ H: (— )-fisetinidol-(+)-catechin

R,^OH, R₂ =H: (— )-robinetinidol-(+)-catechin

R, -' R₂-'OH : (— )-robinetinidol-(- )-gallocatechin

B. 2,3-trans-3,4-cis:2^',3^'-trans-stereoforms F R₂-- H: (— )-fisetinidol-(-i-)-catechin

Table 6

TriflavonoideS

OH

C. Pure trans stereoforms

R OH: (4.8" :4'.6")-( )-robinetinidol-( )-robιnetιnιdol-( I )-gallocatechιn R H [4_,8" :4'.6"]-( )-robιnetιnidol-( • )-ronιnetιnιdol- ( ! ) -catechin

Five iso eric triflavanoides were isolated

Claims

1. Animal feed additive, which comprises a flavanoid containing extract, gained from a plant belonging to the Mimosaceae family, characterized in that it further- more comprises a fragrance and/or taste masking compound or a mixture of such compounds, which mask the fragrance and/or taste of the flavanoid extract, and possibly one or more carriers for the fragrance and/or taste masking compound or the mixture hereof.

2. Additive according to claim 1, characterized in being available in powder or microgranular form or a mixture of these forms.

3. Additive according to claim l or2, characterized in that the flavanoid containing extract is gained from the plant Herba Acacia Catechu Willd. preferably the species Helgatii, Arn. or Acacia mearnsii De Willd (Black Wattle).

4. Additive according to any of the claims 1-3, characterized in that the content of polyhydroxy flavans in the bone dry flavanoid containing Mimosaceae extract constitutes 65-75 % by weight of the total extract mass, preferably 67-72 weight%, and especially about 70 weight%.

5. Additive according to any of the claims 1-4, characterized in that it contains 1-99 weight% flavanoid containing Mimosaceae extract, preferably 5-80 weight%, more preferably 10-50 weight%, most preferably 15-30 weight%, especially about 20 weight%, based upon the total weight of the additive, while the rest is constituted by fragrance and/or taste masking compound, carrier for this and possibly filler and other additions.

6. Additive according to any of the claims 1-5, characterized in that the fra- grance and/or tast masking compound is selected from amongst vanilla, vanilla sugar, whey powder, apple cider powder, and apple vinegar powder or mixtures hereof.

7. Additive according to any of the claims 1-5, characterized in that the smell and/or taste masking compound is brought about in the additive in form of vanilla containing biscuit meal.

8. Animal feed or drink, characterized in comprising a feed or drink, respectively, intended for the pertinent animal and one admixed additive according to any of the claims 1-7, preferably in such weight relationships that the finished manufactured feed or drink, respectively, comprises 0.05 - 10 weight% of the flavanoid containing extract, more preferably 0.1-5 weight%, most preferably 0.5 - 2 weight%, especially about 1 weight%, calculated on the basis of the weight of the total feed or drink mixture.

9. Method for preparation of a feed or drink for prevention of Salmonella ty- phimurium DT 104 in an animal, characterized in that a feed or drink intended for the pertinent animal is admixed to an additive according to one of the claims 1-7, preferably in those in claim 8 stated weight relationships.