CONTROLLED-RELEASE COATED NON-PROTEIN NITROGEN FOOD COMPOSITION AND PROCESS FOR PREPARATION THEREOF

[001] The present invention relates to a controlled-release coated non-protein nitrogen food composition for animal nutrition, particularly to be administered to ruminant animals, which is completely metabolized in the abomasum of the animal. Further, the present invention provides a process for preparing of said composition.

BACKGROUND

[002] The sources of nitrogenous compounds used in cattle feed can be classified as non-protein nitrogen (NPN) and protein nitrogen sources.

[003] Urea and other ammonia forming chemical compounds such as biuret and ammonium sulfate have been widely used in the formulation of diets for beef and milk cattle, especially urea, with the following objectives: (i) reducing costs by replacing part of plant protein sources; and (ii) providing adequate amounts of ruminal degradable protein to better efficiency of fiber digestion and synthesis of microbial protein.

[004] The amount of such compounds, which can safely be used, has been severely limited since these compounds readily form ammonia concentrations in the rumen fluid due to the high hydrolysis rate in the rumen. Ammonia is converted into amino by reaction with carboxylic acids formed from carbohydrate by microbial action in the rumen fluid, and this conversion to carboxylic acid in the rumen is generally unable to keep up with the quick release of ammonia from low substantial soluble sources of non-protein nitrogen (NPN), with nitrogen losses, since there is no synchronism between the carbohydrates fermentation. When absorbed in large amounts, ammonia may exceed the liver detoxification capacity; accumulate in the blood and cause poisoning, which can lead the animal to death.

[005] The high ruminal hydrolysis rate, associated to the animals need to adapt to food with urea, has driven the development of products releasing more slowly this urea in the rumen, but these alternatives are generally more expensive than urea. It is known that complexes of slow urea release can potential reduce toxicity and improve the acceptability and use of urea based concentrates. The gradual release of NH4 allows rumen microorganisms a continuous synthesizing the cellular protein.

[006] Numerous technologies have been developed to synchronize the NPN release rate with carbohydrate degradation rate in the rumen, maximizing the efficiency of microbial protein synthesis. Many of these technologies aimed to the control of release of NPN from urea, which include starea, protection with biuret, liquid urea and calcium chloride, and encapsulating urea with polymeric materials.

[007] Patents Nos. US2560830, US3966998 and US2768895 disclose the use of nonprotein nitrogen (NPN) compounds as constituents supplements in animal nutrition, particularly ruminants.

[008] Patents Nos. US3413118, US3541204, US3925560 and US396699 describe the coating of NPN compounds and metabolizing forms/steps in the rumen and abomasum. However, they do not report about the stability of these layers and the types of products that are applied on the non-protein nitrogen in order to be efficient enough in all metabolism process.

[009] Currently, there are commercially available products under the form of urea or livestock coated urea. However, in both forms, the animal is only able to metabolize from 65% to 75% of its initial volume, bringing the producer to not consider any modification on the urea composition; once your nutrition system only calculates the nitrogen content, considering a high loss of agent efficiency used.

[010] Another disadvantage regarding the encapsulated product is associated with the use of polymeric components of synthetic source, which is not completely metabolized by the animal's digestive system. Despite the polymer components provide proper waterproofing of the urea particle, its ethylenic bonds are difficult to be broken in order to have the urea release and its conversion into ammonium. Thus, it is observed a loss of approximately 30% of the amount administered to the animal.

[011] The present invention provides an improved controlled-release coated nonprotein nitrogen food composition, which is completely metabolized by the animal. The present composition comprises a multilayer coating with natural, animal or vegetal source compounds, being completely metabolizable. Further, the present invention provides a process for preparing thereof. SUMMARY OF THE INVENTION

[012] The instant invention relates to a controlled-release coated non-protein nitrogen food composition for animal nutrition, wherein said composition comprises:

(a) a core, said core comprising 80%-90% wt non-protein nitrogen source (NPN); and

(b) a multilayer coating surrounding said core, in which such coating comprises at least four layers, wherein:

- the first layer comprises 2%-4% wt aldehydes;

- the second layer comprises 2 -4% wt unsaturated fatty acid;

- the third layer comprises l%-3% wt saturated fatty acid; and

- the fourth layer comprises l%-3% wt lubricant agent.

[013] In another embodiment, the present invention refers to a process for preparing said controlled-release coated non-protein nitrogen food composition, wherein said process comprises the following steps:

(c) feeding a fluidized bed equipment in countercurrent with the non-protein nitrogen source (NPN) to form a core;

(d) adding aldehyde compounds in countercurrent to form a first layer;

(e) adding unsaturated fatty acid compounds in countercurrent to form a second layer;

(f) adding saturated fatty acid compounds in countercurrent to form a third layer; and

(g) adding lubricant agent compounds in countercurrent to form a fourth layer. DETAILED DESCRIPTION OF THE INVENTION

[014] The present invention relates to a controlled-release coated non-protein nitrogen food composition wherein said composition is completely metabolized by animals, particularly ruminants. Said composition comprises:

(a) a core, said core comprising 80%-90% wt non-protein nitrogen source (NPN); and

(b) a multilayer coating surrounding said core, in which such coating comprises at least four layers, wherein: - the first layer comprises 2%-4% wt aldehydes;

- the second layer comprises 2%-4% wt unsaturated fatty acid;

- the third layer comprises l%-3% wt saturated fatty acid; and

- the fourth layer comprises l%-3% wt lubricant agent.

[015] The non-protein nitrogen ( PN) source of said core of the present invention is selected from the group consisting of purines, pyrimidines, urea, biuret, thiourea, uric acid, nitrogen glycosides, amides, hidroxialanines, ammonium salts and/or nitrates. Preferably, the non-protein nitrogen (NPN) source is urea.

[016] The composition of the present invention comprises a core which is multilayer coated by insoluble compounds but biodegradable, such as saturated and unsaturated fatty acids, which are deposited on the core constituted by the NPN source so uniform, thereby creating a waterproof barrier.

[017] In the present invention, the multilayer coating system provides the impermeability required for the controlled release and complete metabolism.

[018] Aldehydes which form said first layer are selected from the group consisting of natural alcohols having 2 to 8 carbon atoms.

[019] Unsaturated fatty acids constituting the said second layer are unsaturated fatty acids of vegetable source being selected from the group consisting of oleic acid, linoleic acid, linolenic acid, eicosapentaenoic acid, alpha-linoleic acid, eicosanoic acid, behenic acid, and the like.

[020] Saturated fatty acids constituting said third layer are from vegetable and/or mineral source comprising 18 to 44 carbon atoms, such as, for example, palm oil, coconut oil and stearin of vegetable or animal source. Preferably the mineral source fatty acids are extracted from coal derivatives, natural fossil fuels such as oil and oil shale.

[021] Lubricant agents constituting said fourth layer are vegetable waxes and/or mineral waxes and/or synthetic waxes from petrochemical source. Vegetable waxes are selected from the group consisting of beeswax derivatives, carnauba wax, Montana wax, sugarcane, and the like. Mineral waxes are selected from the group consisting of high molecular weight saturated hydrocarbons. The petrochemical synthetic waxes may be selected from the group consisting of naphtha comprising from 2 to 4 carbon atoms, polyethylene, polypropylene, copolymers thereof and products derived therefrom.

[022] Since components of each layer of the composition of the present invention are natural monomeric, said componets are completely metabolized by the body through the breaking of chemical bonds.

[023] The composition of the present invention has from 0.5 mm to 2.0 mm particle size and 40% to 44% nitrogen release.

[024] In another embodiment, the present invention refers to a process for preparing said controlled-release coated NPN composition, wherein said process comprises the following steps:

c) feeding a fiuidized bed equipment in countercurrent with the non-protein nitrogen ( PN) source to form a core;

d) adding aldehyde compounds in countercurrent to form a first layer;

e) adding unsaturated fatty acid compounds in countercurrent to form a second layer;

f) adding saturated fatty acid compounds in countercurrent to form a third layer; and

g) adding lubricant agent compounds in countercurrent to form a fourth layer.

[025] The process temperature of the present invention is between 30 ° C and 90 ° C; and the process pressure ranges from 0.05 MPa to 0.7 MPa. Preferably, the temperature is between 50 ° C and 90 ° C; and process pressure is between 0.3 MPa to 0.5 MPa.

[026] Throughout the preparation process of said composition of the present invention, the bed pressure is kept constant.

[027] The incorporation and sequencing in fiuidized bed equipment operates on Vander Waals forces, where the orientation of the layers is given by ionic type (hydrogen bonds).

[028] The following examples are presented for illustrative purposes and do not intend to limit the scope of the present invention.

[029] EXAMPLE 1 - Analysis of the complete metabolizing effect [030] The experiment was conducted in the Sector of Dairy Cattle of the Experimental Farm of Gralha Azul (FEGA), located in the campus of Pontificia Universidade Catolica do Parana (PUCPR), located in the municipality of Fazenda Rio Grande in the metropolitan region of Curitiba / PR - BR, by the period of February 1, 2015 to July 1, 2015.

[031] Fifty seven dairy cows were used, of Holstein, red / black and white varieties, with 500 to 700 kg average live weight. The animals were randomly divided into three groups of 19 animals (group 1, group 2 and group 3). These animals were matched according to lactation order, age, milk production and days in milk and distributed in each group in raffle order.

[032] Animals were twice daily milked in a rotary milking parlor, TANDEM model, with twelve milking positions, and equipped with DAIRY PLAN ^® software. The following parameters were evaluated:

• Milk production performance;

• Milk composition (fat content, protein, lactose, total solids and urea nitrogen);

• Somatic cell count (SCC);

• Blood parameters (plasma proteins, albumin and urea nitrogen in plasma);

• Microbial protein absorption Rate by urinary excretion of allantoin; and

• Reproductive parameters related to possible ovarian and uterine changes.

[033] The last milk control data showed the following average on the milk production and udder health:

• Days average lactating = 193.11;

• Age (months) = 36.2;

• Milk production (liters / cow / day) = 32,95; ·% fat milk = 3.61; • % Milk protein = 3.10; ·% total solids = 12.22;

• Somatic cell count (cells / ml) = 351 x 10 ³ ;

• score of body condition = 2.5. [034] Experimental Diets

[035] In this experiment, a crossover design for a full trial period of 63 days was used. Each animals group received a specific diet twice a day after milking, said diet being changed every 21 days.

[036] The diets were composed of three feeds throughout the experimental period, formulated to be isocaloric and isoproteic, taking into account the formulated nutritional levels and obtained by food bromatologic analysis. We tried to keep the same crude protein content estimated in each diet.

[037] Forty five of the fifty seven animals were in the lactation phase, and they were fed for 21 experimentation days in each cycle.

[038] The animals received the following experimental diets:

- Feed A (Control): composed by soybean meal without adding any urea;

- Feed B: composed by a partial substitution of soybean meal by conventional livestock urea meal (with use of 130g livestock urea); and

- Feed C: composed by partial substitution of soybean meal to the composition of the present invention (with use of 136 g composition of the present invention).

[039] Table 1 shows the composition of each diet administered to groups of animals.

[040] Table 1. Composition of experimental diets

Compounds Feed A Feed B Feed C

(kg/v/d) (kg/v/d) (kg/v/d)

Hay 0.370 0.370 0.370

Silage corn 29.00 29.00 29.00

Brewery residue 7.00 7.00 7.00 Soybean meal 1.200

Livestock urea 0.130

Composition of the present invention 0.136

Lactating cows feed 10.81 11.82 11.82 soybean hulls 0.470 0.520 0.520

Wheat bran 2.350 2.590 2.590 corn grain 4.410 4.850 4.850

Soybean meal 3.020 3.240 3.240

Calcitic calcareous 0.260 0.286 0.286

Magnesium oxide 0.050 0.055 0.055 salt 0.050 0.055 0.055 sodium bicarbonate 0.150 0.165 0.165

Premix micro 0.020 0.022 0.022

Oicalcium phosphate 0.010 0.011 0.011

Mastersorb FM 0.020 0.022 0.022

Salt Min. Minerclac 0.250 0.250 0.250

Lactoplus fat 0.150 0.150 0.150 sodium bicarbonate 0.250 0.250 0.250

Total 49.030 48.966 48.972

[041] Table 2 shows the levels of each nutritional diet administered to animals groups.

[042] Table 2 - Nutritional levels of experimental diets

NUTRICIONAL LEVELS Feed A Feed B Feed C

Metabolizable protein (g/v/d) 2432 2314 2448

Ca (g/v/d) 139 144 144 (g/v/d) 81 79 79

Ingestion MS current (kg/v/d) 23.5 23.5 23.5

Ingestion MS predicted (kg/v/d) 23.2 23.2 23.2 PL allowable EM (kg/v/d) 41.2 40.6 40.6

PL allowable PM (kg/v/d) 37.2 34.4 37.5

PDR fed (g/v/d) 2442 2567 2430

PDR balance (g/v/d) 110 246 109

PNDR fed (g/v/d) 1225 1109 1242

PNDR balance (g/v/d) 193 58 211

PB diet (% MS) 15.6 15.6 15.6

PDR diet (% MS) 10.4 10.9 10.3

PNDR diet (% MS) 5.2 4.7 5.3

FDN (% MS) 33.4 33.6 33.6

FDN forage (% MS) 21.7 21.7 21.7

CNF (% MS) 40.4 40.1 40.1

EE (% MS) 4.0 4.1 4.1

Lys (% PM) 6.60 6.65 6.21

Met (% PM) 1.85 1.89 1.78

Lys/Met 3.57 3.52 3.49

Total AAE (g/v/d) 1374 1273 1273

[043] Parameters Evaluated:

[044] All milk composition analyzes were performed in the laboratory of Parana Association of Cattle Breeders of Holstein (APCBRH), located in Curitiba - PR/BR, a member of the Brazilian Milk Quality Network (RBQL).

[045] The urine and blood analyses were carried out by the Laboratory of Clinical Patology, of the Company Animals Hospital Unit of PUCPR in Fazenda Experimental Gralha Azul, located in the municipality of Fazenda Rio Grande - Parana/BR. It was employed the specific commercial kit (DIAGNOSTICA LABTEST S/A), using the endpoint enzymatic colorimetric method in all blood tests.

[046] In the last three days of each cycle, milk production and corrected milk yield for 3.5% fat were individually measured at each milking. [047] Individual milk samples per cycle were collected for analysis: fat (%), protein (%), lactose (%), casein (%) and total solids (%) using the IR method. The milk urea nitrogen (MUN) analysis was performed employing diacetyl monoxime method and flow cytometry was used to count and score the somatic cell.

[048] For plasma urea nitrogen (PUN) analysis, blood samples were collected from six randomized animals per group at each time period of 0, 2, 4, 8 and 10 hours after the morning diet on days 0, 21, 42 and 63.

[049] For analysis of plasma total protein, albumin and creatine, blood samples were collected from ten randomly selected animals per group on days 0, 21, 42 and 63.

[050] Allantoin analysis was carried out by using the colorimetric method described Fujihara et al., ((1987), "The effect of protein infusion on urinary excretion of purine derivates in ruminants nourished by intragastric nutrition"; Journal of Agricultural Science, V. 109, pp. 7-12), and urine samples were collected from ten randomly selected animals per group on days 0, 21, 42 and 63.

[051] We collected a sample of the total mixed diet (TMD) and a sample of the remains of each group on the last day of each cycle. Also it was taken a sample of corn silage cycle, another supplementary roughage (hay Tifton), brewery waste and commercial feed.

[052] Animals were daily monitored by ultrasonography, as the ovarian activity (follicular dynamics, ovulation, the presence of corpus luteum and cysts). The monitoring aims to check possible changes in postpartum animals that meet this period such as, for example, infections and secretory changes in the endometrium.

[053] RESULTS

[054] Table 3 shows the evaluation of milk production where an increase is observed in 10% milk production by about 75% animals.

[055] Table 3 - Evaluation of milk production (%).

Day 1 Day 63

Feed A Feed B Feed C Feed A Feed B Feed C

31.40 23.70 31.70 28.70 24.00 34.20 24.70 35.60 32.90 22.00 27.00 35.20

2730 35.70 35.60 27.00 36.20 29.80

29.10 28.70 39.00 31.70 34.20

40.00 44.20 32.50 37.20 39.30 28.30

23.80 39.20 34.50 24.50 36.60 40.30

34.10 34.50 32.40 38.40 18.90 40.70

32.00 37.20 34.00 28.50 35.30 35.00

42.70 31.80 42.90 44.90 25.90 42.00

33.20 35.60 37.40 31.80 33.20 38.60

28.90 44.20 35.90 31.10 47.90 36.40

38.70 29.50 36.60 37.10 31.90 36.70

28.20 17.20 31.60 25.80 17.80 28.40

28.80 38.20 38.90 25.70 37.50 40.00

31.50 44.90 50.80 32.70 37.80 48.00

27.80 34.50 44.20 12.50 27.30 39.50

47.10 30.50 23.30 48.10 30.80 12.80

42.80 45.20 21.40 43.60 42.40 29.60

30.00 30.40 33.20 36.10 32.40 31.60

[056] Table 4 shows the% fat, where it is observed an increase of 15% fats in about 70% animals.

[057] Table 4 - Evaluation of % fat. D -1 D +69

Feed A Feed B Feed C Feed A Feed B Feed C

(% fat) (% fat) (% fat) (% fat) (% fat) (% fat)

3.06 3.22 2.77

2.68 3.06 1.83

3.09 2.59 3.23

3.40 3.18 2.82

3.99 2.94 3.54

4.25 2.92 3.4

3.53 2.81

3.46 2.96 2.05

2.49 2.38 2.31

2.25 2.1 3.13

2.80 2.66 3.10

2.37 2.93 2.88

2.79 4.22 3.23

2.84 2.68 2.81

3.51 2.19 3.49

3.33 3.05 2.98

2.62 2.97 3.85

2.55 3.26 2.89

2.83 3.08 3.14

2.83 2.00 2.93

3.13 2.70 2.84

2.12 3.59 4.28

3.25 2.82 2.84

3.13 3.62 2.86

3.87 2.87 3.67

3.64 3.06 3.15

3.74 3.07 2.93

3.94 4.22 2.26

4.30 2.57 3.63

4.98 3.21 3.5

2.57 3.08 3.10

1.99 3.55 2.11

3.63 2.30

3.87 2.83 1.97

3.47 2.81 3.60

3.35 2.68 2.49 3.77 3.08 3.08

3.79 3.29 3.14

[058] Table 5 shows the% protein, where we observed an increase of 10% protein at approximately 75% animals.

[059] Table 5 - Evaluation% protein

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Feed A Feed B Feed C Feed A Feed B Feed C

(%protein) (%protein) (%protein) (%protein) (%protein) (%protein)

3.12 3.11 3.22

3.25 3.27 2.93

3.70 3.24 3.25

3.23 3.39 3.07

3.35 3.32 2.99

3.20 3.16 3.12

3.48 3.02 3.20

3.36 2.79 3.12

3.23 2.70 3.19

2.94 2.76 3.13

3.44 3.26 3.07

3.36 3.09 2.88

3.01 3.03 3.02

2.96 3.00 3.19

3.35 3.03 3.17

3.26 2.78 3.09

3.32 3.19 3.37

3.08 3.17 3.13

2.66 2.76 3.36

3.07 2.67 3.14

3.30 2.55 2.91

3.27 2.58 2.71

3.22 2.96 3.34

2.91 2.87 2.74

3.66 3.32 3.59

3.35 3.25 3.29

3.20 3.13 3.00

2.99 2.57 3.06 3.71 3.03 2.94

3.47 2.79 2.78

3.13 2.74 3.02

2.92 2.44 2.92

3.16 2.91

3.01 2.66 3.37

2.94 3.07 3.52

2.63 2.81 3.75

3.44 3.17 3.20

3.26 3.11 3.28

[060] Table 6 shows the% casein, which is observed an increase of 5% casein in about 70% animals.

[061] Table 6 - Assessment of % casein.

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Casein (%} Casein (%)

Feed A Feed B Feed C Feed A Feed B Feed C

2.64 2.64 2.36 2.52 2.70 2.47

2.62 2.76 2.49 2.90 2.98 2.58

2.59 2.56 2.50 2.64 2.58 2.33

2.73 2.23 2.54 2.77 2.57

2.39 2.22 -2.54 2.61 2.17 2.46

2.74 2.49 2.33 2.66 2.57 2.38

2.38 2.41 2.57 2.40 2.36 2.7

2.64 2.22 2.50 2.61 2.61 2.58

2.49 2.57 2.52 2.63 2.48 2.61 2.49 2.13 2.53 1.97 2.32 2.62

2.66 2.02 2.12 2.61 2.07 2.22

2.34 2.27 2.19 2.50 2.33 2.63

2.73 2.64 2.69 2.95 2.75 2.97

2.37 1.99 2.47 2.44 2.38 2.35

2.82 2.21 2.19 2.79 2.41 2.36

2.36 1.89 2.35 2.45 2.08 2.41

2.43 2.13 2.76 2.43 2.21

2.09 2.25 3.12 2.25 2.45 2.72

2.64 2.51 2.68 2.73 2.76 2.25

[062] Table 7 shows the % urea in milk, where it was not observed any significant change.

[063] Table 7 - Evaluation of % urea in milk.

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Urea (mg/dL) Urea (mg/dL)

Feed A Feed B Feed C Feed A Feed B Feed C

14.01 11.63 11.96

15.17 12.45 15.05

14.03 19.43 14.64

10.13 15.26 12.59

13.30 18.71 15.14

10.43 15.30 13.66

15.66 15.17

12.01 12.85 18.45 14.05 13.44 16.07

13.60 14.46 13.29

18.68 21.70 18.36

17.38 17.53 14.96

17.47 11.52 18.89

14.75 12.30 18.65

17.52 14.60 17.57

13.21 17.00 12.77

17.68 18.51 13.03

17.29 15.26 12.91

12.37 14.02 16.65

8.25 12.52 12.00

15.62 13.08 19.16

16.07 9.34 12.01

14.77 15.07 13.17

12.49 8.58 9.54

12.70 17.45 17.84

12.85 17.06 12.69

14.85 15.90 12.91

7.93 7.32 13.25

16.79 13.12 13.91

9.70 10.61 10.14

15.70 15.86 18.74

14.98 10.57 13.08

16.32 44.03

13.07 24.00 16.42

20.07 15.49 19.13

13.02 13.36 24.52

12.54 15.92 11.9

14.28 12.38 11.67

[064] Table 8 shows the % lactose, which is observed an increase of 10% lactose in approximately 75% animals.

[065] Table 8 - Evaluation of % lactose

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Lactose (%) Lactose (%) Feed A Feed B Feed C Feed A Feed B Feed C

4.19 4.65 4.77

4.53 4.38 4.49

4.38 4.64 4.47

4.35 4.70 4.28

4.24 4.55 4.58

4.45 4.60 4.61

4.87 4.27

5.04 3.80 4.37

4.05 4.54 4.38

4.17 4.78 4.51

4.18 4.55 4.80

3.75 4.74 4.90

4.76 4.1 4.69

4.70 4.55 4.58

4.14 4.57 4.44

4.22 4.63 4.72

4.51 4.51 4.62

4.53 4.72 4.69

4.14 4.45 4.65

4.64 4.59 4.71

4.26 4.61 4.40

4.23 4.82 4.39

4.97 4.83 4.59

5.11 4.85 4.30

4.43 4.34 4.26

4.43 3.85 4.23

4.72 4.75 4.39

4.89 4.78 4.34

4.67 4.89 4.57

4.69 4.81 4.64

4.56 4.83 4.61

4.34 4.93 4.93

4.54 4.15

4.81 4.14 4.44

4.86 4.62 4.50

5.03 4.55 4.34

4.81 4.85 4.69

4.60 4.85 4.71 [066] Table 9 shows the evaluation of somatic cells (SCC) in milk, which there were no significant changes.

[067] Table 9 - Evaluation of somatic cells (SCC) in milk.

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SCC (xiOOO/mL) SCC (xlOOO/mL)

Feed A Feed B Feed C Feed A Feed B Feed C

779 143 53 1373 122 52

537 189 100 898 558 116

232 62 17 202 75 27

72 617 123 137 531

85 81 232 133 688 196

1090 40 3 554 272 20

10 246 38 54 339 76

41 171 127 56 791 3436

212 110 141 137 702 138

293 61 388 2564 184 223

303 84 97 422 38 478

67 128 68 75 29 131

75 983 117 164 904 178

8 14 548 80 44 1445 105 10 38 135 31 189

464 33 34 433 53 64

60 229 2217 24 201

5 208 146 51 544 63

110 31 33 98 44 32

[068] Table 10 shows the evaluation of kidney function, which was not observed change.

[069] Table 10 - Analysis of kidney function of ruminants.

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Urine (L/day) Urine (L/day)

Feed A Feed B Feed C Feed A Feed B Feed C

23.68 136.57 62.63 24.58 127.54 28.35

37.66 74.80 115.97 32.72 24.98 43.77

69.45 70.09 37.57 72.99 57.72 21.11

73.75 74.50 36.68 77.44 31.80

32.19 51.13 44.56 22.84 32.42 20.65

53.15 75.85 73.75 49.63 14.16 44.83

41.09 30.11 39.13 20.92 42.10 26.19

65.51 48.17 68.24 25.16 34.04 47.68

161.66 41.27 71.21 75.66 37.69 25.03 43.73 63.84 63.08 38.92 75.98 29.93

42.21 34.08 160.92 44.15 24.51 121.05

31.60 46.26 93.23 23.28 35.13 34.43

73.90 66.87 33.43 31.87 45.38 26.06

86.58 108.23 140.95 74.86 78.56 27.17

31.05 59.70 53.07 46.20 56.88 66.74

{070] Table 11 shows the evaluation of allantoin, where there was no significant change in 75% animals.

[071] Table 11 - Allantoin assessment - nitrogen excretion product by ruminants.

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Alantoin (mg/L) Alantoin (mg/L)

Feed A Feed B Feed C Feed A Feed B Feed C

333.00 264.25 356.75 366.75 360.50 391.75

400.50 426.75 333.00 439.25 496.75 260.50

228.00 340.50 385.50 229.25 226.75 366.75

331.75 346.75 380.50 339.25 378.00

268.00 361.75 420.50 425.50 359.25 494.25

495.50 313.00 453.00 578.00 309.25 458.00

371.75 424.25 489.25 503.00 218.00 494.25

414.25 331.75 380.50 460.50 451.75 340.50 175.50 471.75 268.00 253.00 360.50 443.00

399.25 398.00 444.25 443.00 121.75 495.50

434.25 421.75 140.50 370.50 528.00 129.25

439.25 418.00 269.25 565.50 578.00 451.75

459.25 330.50 386.75 56.75 338.00 413.00

396.75 228.00 491.75 149.25 261.75 436.75

390.50 428.00 405.50 391.75 279.25 486.75

[072] Table 12 shows the evaluation of creatinine - excretory product phosphorylated creatine proportional to the mass muscular-, where there was increase in more than 75% animals.

[073] Table 12 - Creatinine Assessment

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Creatinine (mg/dL) Creatinine (mg/dL)

Feed A Feed B Feed C Feed A Feed B Feed C

72.00 14.00 25.00 72.00 15.50 56.50

47.00 20.00 13.50 55.50 57.00 35.00

20.50 28.00 47.50 20.00 35.00 86.00

24.00 21.50 49.50 20.00 58.00

53.50 31.00 34.00 77.50 50.00 75.00

30.50 19.50 24.00 33.00 107.00 40.50 41.50 52.00 40.50 81.50 38.50 58.50

25.00 32.50 24.00 63.00 46.00 34.00

11.50 40.50 23.00 24.00 42.50 64.00

33.00 27.50 22.00 38.00 22.00 47.00

40.00 46.50 11.00 39.00 62.50 15.00

57.00 36.50 17.00 78.00 49.50 44.50

16.50 20.50 50.50 36.00 31.00 63.50

17.50 14.00 11.50 19.50 19.50 57.00

56.00 28.00 29.50 38.00 28.50 24.00

[074] It can be concluded from the herein-described results that the composition of the present invention was completely metabolized and there was no generation of waste; i.e., both blood and urine samples have not significantly changed.

[075] It should be understood that various modifications and/or alterations may be made without departing from the spirit and scope of the present invention as outlined herein.