The object of the invention is a preparation, being a composition of pancreatic profile enzymes, highly dispersed silicon dioxide, selected organic acids, with the proportions of individual ingredients adapted to age and purpose of the calf, and a method of administering the preparation to calves.

The preparation according to the invention is designed for use in young ruminants, including calves, particularly calves reared for„white meat", i.e. fed essentially only with milk or milk replacement preparations. The effect of using the preparation is an improvement in milk and solid feed assimilation, in particular achieving larger body weight gains in calves. The preparation effectively reduces diarrhea occurrence.

The major problem in calf rearing, especially with calves reared for the so-called white meat, is the digestion and absorption efficiency of all ingredients of milk and milk replacement preparations during the whole fattening period, particularly of fats and proteins.

In case of calf rearing conducted for different purposes (veal, beef, dairy cattle) the essence is to prepare the gastrointestinal tract accustomed to liquid feed (milk and/or milk replacement preparations) to digest and assimilate ingredients of a solid feed.

While there is no threshold feed change in case of calves as it occurs in the case of piglets at the day of weaning, since calf weaning is, in accordance with physiology, a time-extended process, the adjustment (adaptation) of the gastrointestinal tract for complete utilization of solid feed remains however a significant problem.

Hitherto, various treatments were applied, from early administration of solid feed components to pharmacological protection, in particular protection against diarrhea and antibacterial protection of the young animal.

A similar problem exists in calves reared for „white meat", wherein no solid feed is administered, but only milk and/or adapted milk replacement preparations with essential additives (excluding iron).

Research indicates that calves have an innate weaning rhythm and a process associated therewith for the formation of a proventriculuses and for the decrease of exocrine pancreatic functions. Dairy calves produce 1 ml/kg btw of pancreatic juice, and after proventriculus formation 0.2 ml/kg btw (Zabielski R., Pierzynowski, S.G. - Development and regulation of pancreatic juice secretion in cattle. J. Anim. Feed Sci. 10: 25-45, 2001 ). The process of decreasing pancreatic juice secretion in a growing calf is a function of time, does not depend on the feed. Liquid feeds, e.g. milk, which strongly suppress but do not halt proventriculus formation, do not stop the process of decreasing pancreatic juice secretion either, particularly for pancreatic enzymes, such as proteinase, lipase and amylase. The situation is reversed in weaned piglets. Low pancreatic enzyme and pancreatic juice secretion from the milking period is strongly enhanced during weaning period and within 2-3 weeks achieves optimal values for digestion, but this is not always beneficial for obtaining the best growth (Pierzynowski, S.G., Westrom, B., Karlsson, B., Svendsen, L. - Development and regulation of porcine pancreas function - state - of - the - art. Int. J. Pancreatol. 18: 81 -94, 1995).

Microbiological preparations are known, containing pancreatic profile digestive enzymes, as is the possibility for use thereof as pancreatic enzymes in animals being fattened, having a deficiency in their own digestive enzymes (Pierzynowski et al., 2012) and a preparation intended only for piglets during the pre-weaning period (application filed in the Polish Patent Office with the no. P.409972, entitled "Preparation stimulating an improvement in healthiness and weight gains in young mammals, particularly piglets, improving fattening results and a method of administering said preparation"), intended to be used per os.

Various methods of administering therapeutic preparations are known, particularly intravenous, intramuscular, mucosal, rectal, vaginal, as well as per os. Every approach to administration of the preparation, apart from administration per os in water, milk or milk replacement preparation, particularly to calves in first weeks of life, is associated with stress and consequences caused thereby, as well as with the costs of such manipulations.

Unexpectedly, it has been found that administering a mixture of selected pancreatic profile enzymes with highly dispersed silicon dioxide and a mixture of selected organic acids protected in a triglyceride matrix, individually, in drinking water, milk, milk replacement preparations enables active influence on development of ruminant animals, in particular improves nutritional results and cost-effectiveness of animal production, and also eliminates diarrhea which is especially important in case of white meat production. Administering pancreatic profile enzymes was until now considered inadvisable due to suspected existence of a strong mechanism of decreasing secretion of own enzymes after per os administration of pancreatic profile enzymes. This hypothesis has been found to be false in practice. It has been unexpectedly found that pancreatic profile enzymes administered with feed even induce secretion of endogenous enzymes.

An object of a particular embodiment of the invention are four compositions in the form of four sachets intended for specific age groups and purposes of the calves, with a mixture which is ready to use, i.e. to be suspended in water, milk or milk replacement preparation: The first composition is intended for calves within the period of 0-2 weeks of life, regardless of purpose - composition containing pancreatic profile enzymes, highly dispersed silicon dioxide, selected organic acids.

Preferably the composition according to the invention comprises a pancreatic profile enzyme selected from: protease, amylase and lipase, more preferably protease and amylase, particularly preferably protease.

Preferably the composition according to the invention comprises a highly dispersed silicon dioxide having specific surface area of not less than 200 m ² /g.

Preferably the composition according to the invention comprises organic acids selected from sodium salt of butyric acid, fumaric acid, malic acid, citric acid, sorbic acid.

Preferably the composition of the invention comprises a protease in an amount of about 15 mg/day/calf, an amylase in an amount of about 100 mg/day/calf, highly dispersed silicon dioxide in an amount of about 1 .0 g/day/calf, sodium butyrate in an amount of about 150 mg/day/calf, fumaric acid in an amount of about 150 mg/day/calf, malic acid in an amount of about 150 mg/day/calf, citric acid in an amount of about 80 mg/day/calf, sorbic acid in an amount of about 150 mg/day/calf; particularly preferably a protease in an amount of about 6 mg/day/calf, highly dispersed silicon dioxide in an amount of about 1 .0 g/day/calf, fumaric acid in an amount of about 125 mg/day/calf, malic acid in an amount of about 125 mg/day/calf, citric acid in an amount of about 50 mg/day/calf, sorbic acid in an amount of about 100 mg/day/calf.

The second other embodiment of the composition according to the invention is intended for calves within the period of 3-6 weeks of life excluding the calves reared towards white meat - the composition containing pancreatic profile enzymes, highly dispersed silicon dioxide and selected organic acids.

Preferably the composition according to the invention comprises pancreatic profile enzymes selected from: protease, amylase and lipase, preferably protease and lipase, particularly preferably protease, amylase and lipase.

Preferably the composition according to the invention comprises a highly dispersed silicon dioxide having specific surface area of not less than 200 m ² /g.

Preferably the composition according to the invention comprises organic acids selected from sodium salt of butyric acid, fumaric acid, malic acid, citric acid, sorbic acid.

Preferably the composition of the invention comprises a protease in an amount of about 20 mg/day/calf, a lipase in an amount of about 100 mg/day/calf, highly dispersed silicon dioxide in an amount of about 2.0 g/day/calf, sodium butyrate in an amount of about 200 mg/day/calf, fumaric acid in an amount of about 200 mg/day/calf, malic acid in an amount of about 200 mg/day/calf, citric acid in an amount of about 100 mg/day/calf, sorbic acid in an amount of about 150 mg/day/calf; particularly preferably a protease in an amount of about 12 mg/day/calf, a lipase in an amount of about 12 mg/day/calf, an amylase in an amount of about 12 mg/day/calf, highly dispersed silicon dioxide in an amount of about 1.5 g/day/calf, fumaric acid in an amount of about 185 mg/day/calf, malic acid in an amount of about 185 mg/day/calf, citric acid in an amount of about 75 mg/day/calf, sorbic acid in an amount of about 150 mg/day/calf.

The third other embodiment of the composition according to the invention is intended exclusively for calves reared towards white meat within the period of 3 ^rd -8 ^th week. It is a composition comprising a mixture of pancreatic profile enzymes, highly dispersed silicon dioxide and selected organic acids.

Preferably the composition according to the invention comprises pancreatic profile enzymes selected from: protease, amylase and lipase, preferably protease and lipase, particularly preferably protease, amylase and lipase.

Preferably the composition according to the invention comprises a highly dispersed silicon dioxide having specific surface area of not less than 200 m ² /g.

Preferably the composition according to the invention comprises organic acids selected from sodium salt of butyric acid, fumaric acid, malic acid, citric acid, sorbic acid.

Preferably the composition of the invention comprises a protease in an amount of about 25 mg/day/calf, a lipase in an amount of about 150 mg/day/calf, highly dispersed silicon dioxide in an amount of about 2.5 g/day/calf, sodium butyrate in an amount of about 250 mg/day/calf, fumaric acid in an amount of about 250 mg/day/calf, malic acid in an amount of about 200 mg/day/calf, citric acid in an amount of about 100 mg/day/calf, sorbic acid in an amount of about 200 mg/day/calf; particularly preferably a protease in an amount of about 12 mg/day/calf, a lipase in an amount of about 30 mg/day/calf, an amylase in an amount of about 12 mg/day/calf, highly dispersed silicon dioxide in an amount of about 2.0 g/day/calf, fumaric acid in an amount of about 185 mg/day/calf, malic acid in an amount of about 185 mg/day/calf, citric acid in an amount of about 75 mg/day/calf, sorbic acid in an amount of about 150 mg/day/calf.

The fourth other embodiment of the composition according to the invention is intended exclusively for calves reared towards„white meat" within the period from 9 ^th week till the end of fattening. It is a composition comprising a mixture of pancreatic profile enzymes, highly dispersed silicon dioxide and selected organic acids.

Preferably the composition according to the invention comprises pancreatic profile enzymes selected from: protease, amylase and lipase, preferably protease and lipase, particularly preferably protease, amylase and lipase.

Preferably the composition according to the invention comprises a highly dispersed silicon dioxide having specific surface area of not less than 200 m ² /g.

Preferably the composition according to the invention comprises organic acids selected from sodium salt of butyric acid, fumaric acid, malic acid, citric acid, sorbic acid.

Preferably the composition of the invention comprises a protease in an amount of about 40 mg/day/calf, a lipase in an amount of about 250 mg/day/calf, highly dispersed silicon dioxide in an amount of about 2.5 g/day/calf, sodium butyrate in an amount of about 400 mg/day/calf, fumaric acid in an amount of about 400 mg/day/calf, malic acid in an amount of about 400 mg/day/calf, citric acid in an amount of about 200 mg/day/calf, sorbic acid in an amount of about 300 mg/day/calf; particularly preferably a protease in an amount of about 18 mg/day/calf, a lipase in an amount of about 45 mg/day/calf, an amylase in an amount of about 18 mg/day/calf, highly dispersed silicon dioxide in an amount of about 3.0 g/day/calf, fumaric acid in an amount of about 277.5 mg/day/calf, malic acid in an amount of about 277.5 mg/day/calf, citric acid in an amount of about 1 12.5 mg/day/calf, sorbic acid in an amount of about 225 mg/day/calf.

A further object of the invention is the use of the above defined composition according to the invention as an additive to milk, milk replacement preparations or water.

Preferably, composition no. 1 (preparation no. 1) according to the invention as defined above is used daily during the period from birth until the end of the 2 ^nd week of life, preferably within 2-14 days of life, particularly preferably from 4 ^th to 14 ^th day of life regardless of the purpose of the calves (veal, beef, dairy cows, white meat). Preparation no. 1 (one package) is suspended in a small amount of milk, milk replacement preparation or water and administered individually to each calf separately, preferably before feeding.

Preferably, composition no. 2 (preparation no. 2) according to the invention as defined above is used daily during the period from 3 ^rd to 6 ^th week of life and is intended for rearing calves for veal, beef and dairy cows. Preparation no. 2 (one package) is suspended in a small amount of milk, milk replacement preparation or water and administered individually to each calf separately, preferably before feeding. Preferably, composition no. 3 (preparation no. 3) according to the invention as defined above is used daily during the period from 2 ^nd to 8 ^th week of life, being particularly intended for calves for white meat. Preparation no. 3 (one package) is suspended in a small amount of milk, milk replacement preparation or water and administered individually to each calf separately, preferably before feeding.

Preferably, composition no. 4 (preparation no. 4) according to the invention as defined above is used daily during the period from the 9 ^th week of life till the end of fattening, being particularly intended for calves for „white meat". Preparation no. 3 (one package) is suspended in a small amount of milk, milk replacement preparation or water and administered individually to each calf separately, preferably before feeding.

The preparation according to the invention, stimulating digestive and absorption processes, growth, reducing diarrhea, increasing immunity of the animal, is characterized by the fact that it is a mixture of pancreatic profile enzymes, particularly protease and/or lipase and/or amylase with highly dispersed silicon dioxide, with selected organic acids, wherein relative proportions of the ingredients and daily doses depend on age and purpose of the calf.

For determination of the content of preparation ingredients that are stimulating digestive and absorption processes, growth, reducing diarrhea, increasing immunity of the animal, routine analytical methods are applied.

The term "pancreatic profile enzymes" as used herein refers to digestive enzymes produced by the pancreas or known enzymes produced by microorganisms having amylolytic, proteolytic or lipolytic activity similar to the activity of animal pancreatic enzymes. In particular, examples of such enzymes are amylase from Aspergillus oryzae, protease (caseinolytic) from Aspergillus melleus or lipase from Burholderia cepacia.

The generally accepted measure of enzyme activity is the amount of transformed substrate or the formed product in a unit of time.

An exemplary amylolytic enzyme is oamylase (EC 3.2.1 .1), which hydrolyzes an a-1 ,4-D-glycoside bond in polysaccharides comprising three or more D-glucose units linked with an a-1 ,4-D-glycoside bond. The substrate for performing the activity assay is starch. Measurement is done by titrating iodine released under assay conditions.

The term "U" or "unit" refers to enzymatic activity. For example, one USP unit of amylase activity is the amount of enzyme degrading starch with an initial rate of 0.16 μΕς. USP units are not equivalent to the units established in European Pharmacopea (ph.eur.) and need to be converted in the following way (Scharpe et al., 1997): For amylase: 1 ph.eur. unit = 1 BP unit = 1 FIP unit = 4.15 USP units. Activities can be determined with a method presented in European Pharmacopea, which is generally identical with the methodology presented by FIP (Lauwers and Scharpe, 1997). In the known method, α-amylase releases reducing groups from starch, the groups reacting in alkaline solution with iodine present in excess under assay conditions. The amount of iodine that did not react in the reaction is determined by titrating with thiosulfate. Amylolytic activity is determined by comparing the rate at which a pancreatic lyophilisate suspension hydrolyzes a starch substrate solution with the rate at which the standard pancreatic lyophilisate BRP suspension (amylase and lipase) hydrolyzes the same substrate in the same conditions.

An exemplary proteolytic enzyme is a protease, such as pancreatin or pancreatin-like enzymes, which hydrolyze casein to short peptides. For activity determination, under assay conditions the amount of peptides that cannot be precipitated with trichloroacetic acid is determined spectrophotometrically for wavelength of λ 275 nm. The total proteolytic activity of a protease is determined by comparing the amount of non-precipitated peptides released per minute from a casein solution with the amount of such peptides released from the same substrate by the standard pancreatic lyophilisate BRP suspension under the same assay conditions. One USP unit of protease activity is equivalent to the amount of protease that hydrolyzes casein with such rate that in one minute peptides which are not precipitated by trichloroacetic acid, are released in an amount that results in the same absorbance at wavelength of λ 280 nm as a 15 nmol tyrosine solution. USP units are not equivalent to the units established in European Pharmacopea (ph.eur.) and need to be converted in the following way (Scharpe et al., 1997): For protease: 1 ph.eur. unit = 1 BP unit = 1 FIP unit = 62.5 USP units.

Lipase activity can be determined with the method provided in European Pharmacopea, which is generally identical with the methodology shown by FIP (Lauwers and Scharpe, 1997).

The basic method of determination of lipase activity is the titration method, wherein the concentration of fatty acids being released by lipase from an olive oil emulsion is determined with sodium hydroxide.

Lipolytic activity is determined by comparing the rate at which an enzyme suspension hydrolyzes a substrate of olive oil emulsion with the rate at which a suspension of the pancreas (amylase and lipase) BRP powder hydrolyzes the same substrate under the same conditions. For example: 1 USP unit of lipase activity is contained in the amount of pancreatin that releases 1 .0 μEq of acid per minute at pH 9.0 and +37°C. USP units are direct equivalents to ph.eur. units. For lipase: 1 ph.eur. unit = 1 BP unit = 1 FIP unit = 1 USP unit.

The term "highly dispersed silicon dioxide" as used herein refers to dispersion at which the surface area of silicon dioxide is not less than 200 m ² /g.

Organic acids are measured with a high-performance liquid chromatography (HPLC) method.

The aim and scope of application: determination of organic acids (citric acid, formic acid, fumaric acid, malice acid, sodium butyrate and lactic acid) with HPLC in a powdery, granulated or protected in triglyceride matrices feed additive.

The principle: citric acid, formic acid, malic acid, sodium butyrate and lactic acid are measured quantitatively, after reconstitution in water, with HPLC using a UV detector - detection at wavelength of 214 and 254 nm.

Sorbic acid is measured after dissolution in methanol with a HPLC method using a UV detector - wavelength of 254 nm.

The term "white meat" as used herein refers to white calf meat. White veal must originate from calves slaughtered up to the 8 ^th month of life, being designated as veal but with white color. Calves must be fed according to a program based on milk feeds with partial use of soy proteins.

The term "gastro-intestinal disease" as used herein refers to a disease of gastro-intestinal tract, such as a disease of the esophagus, stomach, first, second and third part of the duodenum, jejunum, ileum, ileocecal valve, colon, sigmoid and rectum. Gastro-intestinal diseases, in particular gastro-intestinal diseases characterized with diarrhea, may be associated with various etiological factors. Causative factors may be, but are not limited to, bacteria, viruses, rotaviruses, protozoa, stress factors, nutrition etc. The main bacteria causing gastro-intestinal diseases are Escherichia coli (E. coli) and members of the Clostridium, Lawsonia and Brachyspira genera. The main viruses causing gastro-intestinal diseases are rotaviruses, coronaviruses and infectious viruses of the gastro-intestinal tract. Apart from bacterial and viral infections, gastro-intestinal diseases may be caused by various alterations, such as stress, which may be associated with irregular feed intake, feed structure, animal hygiene and zoohygienic conditions of the space where the animals are accommodated (humidity, temperature, air movement) and with inadequate space for feeding in the pen. Gastro-intestinal disease may also be caused by (early) weaning. Insufficient provision of colostrum to a newborn calf. In fact, it has been observed that in the body of an early-weaned one multiple changes occur, such as morphological and functional changes in small intestine, particularly in intestinal epithelia, changes in colonization of small intestine with prevalence of E. coli and impairment of immunological system. Non-infectious stress factors which are associated with development of a gastro-intestinal disease, for example in calves, may be, but are not limited to, age of the calves at the moment of weaning and the abrupt change in feed from milk providing the calves with immunoglobulin.

Thus, in particular embodiments of the invention the gastro-intestinal disease is an intestinal disease. The term "intestinal disease" as used herein relates to an intestine or is associated with intestine dysfunction. The gastro-intestinal disease or intestinal disease may be characterized with diarrhea. The term "diarrhea" as used herein refers to a condition wherein there are three or more loose or liquid stools or bowel movements per day.

The term "depriving of mother's milk" or "weaning" as used herein refers to introducing an infant or a young mammal to what will be the adult feed and discontinuation of providing of mother's milk or a bottled substitute. An infant is considered "weaned" when no longer receiving any mother's milk or bottled substitute. In particular embodiments, a composition enriched with a glycoprotein or nutritional supplements or feeds according to the present invention are designed for use in treatment or prevention of gastro-intestinal diseases in weaned animals, more particularly in weaned livestock animals. The inventors have experimentally found that the composition from 1 to 4 according to the present invention show a positive effect on treatment of gastro-intestinal diseases. In particular, the inventors have found that the compositions according to the present invention show a positive effect on prevention and treatment of diarrhea. Accordingly, it is expected that the compositions according to the present invention are useful for treatment or prevention of gastro-intestinal diseases, more particularly intestinal diseases and most particularly gastro-intestinal diseases and/or intestinal diseases characterized with diarrhea, in a subject in need thereof. The compositions according to the present invention are furthermore provided for use in treatment and prevention of diarrhea.

The inventors have also experimentally demonstrated that the compositions from 1 to 4 according to the present invention show a positive effect on digestive processes and feed absorption, manifesting in increased weight gains in calves with a more advantageous feed consumption coefficient, which is particularly beneficial in the case of rearing calves towards "white meat".

The invention solves the issue of adapting the gastro-intestinal tract of a calf for effective feed intake, particularly in the case of production of the so-called white meat, that is it minimizes the problems associated with feeding with milk or milk replacement preparations only. The essence of the invention involves use of the above defined composition in order to aid and regulate digestive processes in calves and to improve the absorption capability for nutritional ingredients of feed, particularly in the case of feeding with milk or milk replacement preparations only. As a result, the possibility of effects accompanying calf rearing, such as diarrhea, poor gains, unsatisfactory feed utilization, lack of appetite, susceptibility to infections, is minimized.

Example 1 . Preparation no. 1 being a composition of protease, highly dispersed silicon dioxide, fumaric acid, malic acid, citric and sorbic acid.

The contents of 1 package of the preparation:

6 mg of protease DS (Amamo Enzyme Inc or a pancreatic profile protease from another manufacturer) with activity of not less than 22,000, - ph.eur. units/g, i.e. about 132 ph.eur. units of protease in total,

1000 mg of highly dispersed silicon dioxide (Aerosil®A300F) of A300 Pharma quality or equivalent (see characteristics of A300F from Aerosil),

125 mg of fumaric acid of FCC or higher

125 mg of malic acid of FCC quality or higher

50 mg of citric acid of FCC quality or higher

100 mg of sorbic acid of FCC quality or higher

Use: 1 sachet per day, suspended in milk, water, milk replacement preparation, individually for a calf, since birth until the 14 ^th day of life, regardless of the purpose of the calf.

Production of the preparation:

Previously weighed amounts of highly dispersed silicon dioxide, protease, fumaric acid, malic acid, citric acid and sorbic acid are added to a prepared mixer/homogenizer. The mixture is homogenized by activating the mixing systems.

After analysis, the final product is packed into sachets labeled accordingly, the sachets are packed into bulk boxes and these are palletized. After the final quality control the product is routed for sale.

Example 2. Preparation no. 2 being a composition of protease, lipase, amylase, highly dispersed silicon dioxide, fumaric acid, malic acid, citric and sorbic acid.

The contents of 1 package of the preparation:

12 mg of protease DS (Amamo Enzyme Inc or a pancreatic profile protease from another manufacturer) with activity of not less than 22,000, - ph.eur. units/g, i.e. about 264 ph.eur. units of protease in total,

12 mg of lipase DS (Amamo Enzyme Inc or a pancreatic profile lipase from another manufacturer) with activity of not less than 23,000, - ph.eur. units/g, i.e. about 276 ph.eur. units of lipase in total, another manufacturer) with activity of not less than 24,000, - ph.eur. units/g, i.e. about 288 ph.eur. units of amylase in total,

1500 mg of highly dispersed silicon dioxide (Aerosil®A300F) of A300 Pharma quality or equivalent (see characteristics of A300F from Aerosil),

185 mg of fumaric acid of FCC or higher

185 mg of malic acid of FCC quality or higher

75 mg of citric acid of FCC quality or higher

150 mg of sorbic acid of FCC quality or higher

Use: 1 sachet per day, suspended in milk, water, milk replacement preparation, individually for a calf, from the 15 ^th to the 42 ^nd day of life (week 3-6), excluding calves reared for "white meat".

Production of the preparation:

Previously weighed amounts of highly dispersed silicon dioxide, protease, lipase, amylase, fumaric acid, malic acid, citric acid and sorbic acid are added to a prepared mixer/homogenizer. The mixture is homogenized by activating the mixing systems.

After analysis, the final product is packed into sachets labeled accordingly, the sachets are packed into bulk boxes and these are palletized. After the final quality control the product is routed for sale.

Example 3. Preparation no. 3 being a composition of protease, lipase, amylase, highly dispersed silicon dioxide, fumaric acid, malic acid, citric and sorbic acid.

The contents of 1 package of the preparation:

12 mg of protease DS (Amamo Enzyme Inc or a pancreatic profile protease from another manufacturer) with activity of not less than 22,000, - ph.eur. units/g, i.e. about 264 ph.eur. units of protease in total,

30 mg of lipase DS (Amamo Enzyme Inc or a pancreatic profile lipase from another manufacturer) with activity of not less than 23,000, - ph.eur. units/g, i.e. about 690 ph.eur. units of lipase in total,

12 mg of amylase DS (Amamo Enzyme Inc or a pancreatic profile amylase from another manufacturer) with activity of not less than 24,000, - ph.eur. units/g, i.e. about 288 ph.eur. units of amylase in total,

2000 mg of highly dispersed silicon dioxide (Aerosil®A300F) of A300 Pharma quality or equivalent (see characteristics of A300F from Aerosil),

185 mg of fumaric acid of FCC or higher 185 mg of malic acid of FCC quality or higher

75 mg of citric acid of FCC quality or higher

150 mg of sorbic acid of FCC quality or higher

Use: 1 sachet per day, suspended in milk, water, milk replacement preparation, individually for a calf, from the 15 ^th to the 54 ^th day of life (week 3-8), exclusively for calves reared for "white meat".

Production of the preparation:

Previously weighed amounts of highly dispersed silicon dioxide, protease, lipase, amylase, fumaric acid, malic acid, citric acid and sorbic acid are added to a prepared mixer/homogenizer. The mixture is homogenized by activating the mixing systems.

After analysis, the final product is packed into sachets labeled accordingly, the sachets are packed into bulk boxes and these are palletized. After the final quality control the product is routed for sale.

Example 4. Preparation no. 4 being a composition of protease, lipase, amylase, highly dispersed silicon dioxide, fumaric acid, malic acid, citric and sorbic acid.

The contents of 1 package of the preparation:

18 mg of protease DS (Amamo Enzyme Inc or a pancreatic profile protease from another manufacturer) with activity of not less than 22,000, - ph.eur. units/g, i.e. about 396 ph.eur. units of protease in total,

45 mg of lipase DS (Amamo Enzyme Inc or a pancreatic profile lipase from another manufacturer) with activity of not less than 23,000, - ph.eur. units/g, i.e. about 1035 ph.eur. units of lipase in total,

18 mg of amylase DS (Amamo Enzyme Inc or a pancreatic profile amylase from another manufacturer) with activity of not less than 24,000, - ph.eur. units/g, i.e. about 432 ph.eur. units of amylase in total,

3000 mg of highly dispersed silicon dioxide (Aerosil®A300F) of A300 Pharma quality or equivalent (see characteristics of A300F from Aerosil),

277.5 mg of fumaric acid of FCC or higher

277.5 mg of malic acid of FCC quality or higher

1 12.5 mg of citric acid of FCC quality or higher

225 mg of sorbic acid of FCC quality or higher Use: 1 sachet per day, suspended in milk, water, milk replacement preparation, individually for a calf, from the 43 ^rd day of life until slaughter (week 9-16), exclusively for calves reared for "white meat".

Production of the preparation:

Previously weighed amounts of highly dispersed silicon dioxide, protease, lipase, amylase, fumaric acid, malic acid, citric acid and sorbic acid are added to a prepared mixer/homogenizer. The mixture is homogenized by activating the mixing systems.

After analysis, the final product is packed into sachets labeled accordingly, the sachets are packed into bulk boxes and these are palletized. After the final quality control the product is routed for sale.

DESCRIPTION OF EXPERIMENT NO. 1 - calves intended for "white meat".

A research experiment was conducted as commissioned by the inventors. The experiment included 20 calves, mixed cb x Hf breeds (50-75% of Hf genes) weaned at the moment of termination of colostrum feeding (4 ^th day of life). The animals being born were assigned to 2 groups: control and experimental. Each group had 10 calves kept in individual pens. At the start of the experiment the animals were weighed, subsequent measurements of body weight were in 10-day intervals.

Calf feed was based on milk administered twice a day until the 7 ^th day, and from the 7 ^th day of life a concentrated blend was introduced to the diet (control group).

The calves from the experimental group were administered whole milk as exclusive feed during the whole rearing period, as well as three experimental preparations added to the milk, in their respective periods: preparation 1 from the 4 ^th day till the 2 ^nd week of life, subsequently the calves were administered preparation no. 3 from the 3 ^rd until the end of the 8 ^th week of life, whereupon from the 9 ^th week of life they were administered preparation 4 until the end of the experiment in the 17 ^th week of life (age 120 days).

Animals from the control group were fed individually with daily weighing of the feeds and controlling the amount of feed refusal, while the animals from the experimental group did not receive solid feed and were fed individually with milk including additives. During the experiment, the number of days with diarrhea, the course thereof (clinical observations) were reported on a health chart, health condition was determined. Body weight gains and feed consumption were determined. At the beginning of the experiment (4 ^th day of life), as well as on the 60 ^th day and at the end of the experiment (day 120) blood was collected from external jugular vein in order to determine morphological (hematocrit, hemoglobin content, red and white cell count) and biochemical indicators in blood (α, β, γ immunoglobulin levels).

Statistical calculations were done using univariate ANOVA. The significance of differences between the experimental groups was estimated with the Tukey's test using Statgraphics Plus 5.1 software (2001 ). Differences were considered statistically significant at the significance level P<0.05.

Results

Table 1 . Results of calf rearing

Values designated with the same letters differ significantly from one another - capital letters p<0.01 , small letters p<0.05 Table 2. Biochemical and hematological indicators in calf blood serum

p <0.05 Conclusions

The obtained results demonstrated positive effect of administering a milk additive of preparations comprising pancreatic profile enzymes, highly dispersed silicon dioxide, and selected organic acids on health indicators and rearing of calves since birth till 120 days of age.

In the experimental group a significant reduction of diarrhea occurrence was observed. Good body weight gains were reported in both groups, low feed consumption per 1 kg of weight gain was observed in the control group (2.52 ± 0.14 kg/kg body weight).

In the experimental group a reduction in red blood cell parameters was observed in morphological blood analysis, including a decrease in red blood cell count and hemoglobin levels in comparison to the control group animals. Significantly increased γ group immunoglobulin concentrations were found in blood plasma of the experimental group animals.

The application of preparations comprising pancreatic profile enzymes, highly dispersed silicon dioxide, and selected organic acids in calf rearing was found to be effective means for prevention and amelioration of results of diarrhea in calves, had a positive effect on rearing parameters (body weight gains) and feed consumption.

DESCRIPTION OF EXPERIMENT NO. 2 - calves intended for purposes other than "white meat".

The experiment was conducted on 20 calves, mixed cb x Hf breeds (50-75% of Hf genes) weaned at the moment of termination of colostrum feeding (4 ^th day of life). The animals being born were assigned to 2 experimental groups. Each group had 10 calves kept in individual pens with automated troughs, on an openwork plastic flooring. At the start of the experiment the animals were weighed, subsequent measurements of body weight were in 10-day intervals, and on the day ending he experiment, i.e. on the 42 ^nd day of life, the test being conducted within the period from the 1 ^st to 42 ^nd day of life of the calves.

The calves were fed with whole milk according to the IZ PIB-INRA, 2009 standards. Calf feeding was based on whole milk administered twice a day until the 7 ^th day, and from the 4 ^th day of life a concentrated blend was introduced to the diet (control group and experimental group). The animals of both groups were administered concentrated blend ad libitum and meadow hay in the amount of 0.10 kg/day during the period of milk feeding to 0.30 kg/day after its completion. Nutrient content in the concentrated blend in 1 kg DM was 206 g of total protein, 24 g of crude fat, 59 g of crude fiber and 32 g of crude ash, respectively. The calves from the experimental group were administered identical feeding during the whole experimental period, supplemented by the addition of three preparations administered in milk in in their respective periods: preparation 1 from the 4 ^th day till the 2 ^nd week of life, subsequently the calves were administered preparation no. 2 from the 3 ^rd until the end of the 6 ^th week of life.

During the experiment, the number of days with diarrhea, the course thereof (clinical observations) were reported on a health chart, health condition was determined. Body weight gains and feed consumption were determined.

Values designated with the same letters differ significantly from one another - capital letters p<0.01 , small letters p<0.05 Discussion and conclusions

The obtained results demonstrated positive effect of administering a milk additive of preparations comprising pancreatic profile enzymes, highly dispersed silicon dioxide, selected organic acids on health indicators and rearing of calves since birth till 42 days of age.

In the conducted experiment, high and balanced body weight gains were observed for calves, on the 42 ^nd day of life significantly higher body weight gains for experimental group calves were reported (p≥0.01 ). Calves of both nutritional groups were characterized by a high feed consumption coefficient, which indicates an adequate digestive process and good health state of the animels.

The applied additives and system of calf weaning caused sporadic occurrences of diarrhea, however in the second part of the experiment, within the period between the 15 ^th and 42 ^nd day of life a reduction in prevalence of diarrhea in experimental group calves by 21 .5% (28 vs 22) was observed.

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